Pyrrolobenzodiazepines

ABSTRACT

A compound with the formula I, wherein R 2  is of formula II, where A is a C 5-7  aryl group, X is selected from the group consisting of: OH, SH, CO 2 H, COH, N═C═O, NHNH 2 , CONHNH 2 , (III), (IV), and NHR N , wherein R N  is selected from H and C 1-4  alkyl, and either: (i) Q 1  is a single bond, and Q 2  is selected from a single bond and —Z—(CH 2 ) n —, where Z is selected from a single bond, O, S and NH and n is from 1 to 3; or (ii) Q 1  is —CH═CH—, and Q 2  is a single bond; and its conjugates.

CROSS REFERENCE TO RELATED APPLICATIONS

The present application is a national phase application under 35 U.S.C.§ 371 of PCT International Application No. PCT/EP2017/051600, filed Jan.26, 2017, which claims priority to Great Britain Application No.1601431.8, filed Jan. 26, 2016, each of which are hereby incorporated byreference in its entirety.

INCORPORATION-BY-REFERENCE OF MATERIAL SUBMITTED ELECTRONICALLY

Incorporated by reference in its entirety herein is a computer-readablenucleotide/amino acid sequence listing submitted concurrently herewithand identified as follows: One 47,858 Byte ASCII (Text) file named“36774-US-1-PCT_ST25.TXT,” created on Jun. 27, 2019.

The present invention relates to pyrrolobenzodiazepines (PBDs), inparticular pyrrolobenzodiazepine dimers having a C2-C3 double bond andan aryl group at the C2 position on one monomer unit, and an exo doublebond at the C2 position on the other monomer unit.

BACKGROUND TO THE INVENTION

Some pyrrolobenzodiazepines (PBDs) have the ability to recognise andbond to specific sequences of DNA; the preferred sequence is PuGPu. Thefirst PBD antitumour antibiotic, anthramycin, was discovered in 1965(Leimgruber, et al., J. Am. Chem. Soc., 87, 5793-5795 (1965);Leimgruber, et al., J. Am. Chem. Soc., 87, 5791-5793 (1965)). Sincethen, a number of naturally occurring PBDs have been reported, and over10 synthetic routes have been developed to a variety of analogues(Thurston, et al., Chem. Rev. 1994, 433-465 (1994)). Family membersinclude abbeymycin (Hochlowski, et al., J. Antibiotics, 40, 145-148(1987)), chicamycin (Konishi, et al., J. Antibiotics, 37, 200-206(1984)), DC-81 (Japanese Patent 58-180 487; Thurston, et al., Chem.Brit., 26, 767-772 (1990); Bose, et al., Tetrahedron, 48, 751-758(1992)), mazethramycin (Kuminoto, et al., J. Antibiotics, 33, 665-667(1980)), neothramycins A and B (Takeuchi, et al., J. Antibiotics, 29,93-96 (1976)), porothramycin (Tsunakawa, et al., J. Antibiotics, 41,1366-1373 (1988)), prothracarcin (Shimizu, et al, J. Antibiotics, 29,2492-2503 (1982); Langley and Thurston, J. Org. Chem., 52, 91-97(1987)), sibanomicin (DC-102)(Hara, et al., J. Antibiotics, 41, 702-704(1988); Itoh, et al., J. Antibiotics, 41, 1281-1284 (1988)), sibiromycin(Leber, et al., J. Am. Chem. Soc., 110, 2992-2993 (1988)) and tomamycin(Arima, et al., J. Antibiotics, 25, 437-444 (1972)). PBDs are of thegeneral structure:

They differ in the number, type and position of substituents, in boththeir aromatic A rings and pyrrolo C rings, and in the degree ofsaturation of the C ring. In the B-ring there is either an imine (N═C),a carbinolamine(NH—CH(OH)), or a carbinolamine methyl ether (NH—CH(OMe))at the N10-C11 position which is the electrophilic centre responsiblefor alkylating DNA. All of the known natural products have an(S)-configuration at the chiral C11a position which provides them with aright-handed twist when viewed from the C ring towards the A ring. Thisgives them the appropriate three-dimensional shape for isohelicity withthe minor groove of B-form DNA, leading to a snug fit at the bindingsite (Kohn, In Antibiotics III. Springer-Verlag, New York, pp. 3-11(1975); Hurley and Needham-VanDevanter, Acc. Chem. Res., 19, 230-237(1986)). Their ability to form an adduct in the minor groove, enablesthem to interfere with DNA processing, hence their use as antitumouragents.

It has been previously disclosed that the biological activity of thismolecules can be potentiated by joining two PBD units together throughtheir C8/C′-hydroxyl functionalities via a flexible alkylene linker(Bose, D. S., et al., J. Am. Chem. Soc., 114, 4939-4941 (1992);Thurston, D. E., et al., J. Org. Chem., 61, 8141-8147 (1996)). The PBDdimers are thought to form sequence-selective DNA lesions such as thepalindromic 5′-Pu-GATC-Py-3′ interstrand cross-link (Smellie, M., etal., Biochemistry, 42, 8232-8239 (2003); Martin, C., et al.,Biochemistry, 44, 4135-4147) which is thought to be mainly responsiblefor their biological activity. One example of a PBD dimer is SG2000(SJG-136):

(Gregson, S., et al., J. Med. Chem., 44, 737-748 (2001); Alley, M. C.,et al., Cancer Research, 64, 6700-6706 (2004); Hartley, J. A., et al.,Cancer Research, 64, 6693-6699 (2004)).

Dimeric PBD compounds bearing C2 aryl substituents, such as SG2202(ZC-207), are disclosed in WO 2005/085251:

and in WO2006/111759, bisulphites of such PBD compounds, for exampleSG2285 (ZC-423):

These compounds have been shown to be highly useful cytotoxic agents(Howard, P. W., et al., Bioorg. Med. Chem. (2009), doi:10.1016/j.bmc1.2009.09.012).

Due to the manner in which these highly potent compounds act incross-linking DNA, these molecules have been made symmetrically. Thisprovides for straightforward synthesis, either by constructing the PBDmoieties simultaneously having already formed the dimer linkage, or byreacting already constructed PBD moieties with the dimer linking group.

WO 2010/043880 and WO 2011/130613 disclose dimeric PBDs similar to thoseabove which are not symmetrical due to a group on one of the C2 arylgroups which can be joined to a linker to a cell binding agent, such asan antibody.

WO 2011/130616 discloses similar PBD dimer conjugates, where the PBDmonomer which is not linked to the cell binding agent has a double bondbetween C2 and C3 but a non-aromatic C2 substituent.

WO 2013/053873 discloses PBD dimer conjugates, where the PBD monomerlinked to the cell binding agent has a double bond between C2 and C3 andis linked via a propenyl group, and the PBD monomer which is not linkedto the cell binding agent has a double bond between C2 and C3 and anaromatic or non-aromatic C2 substituent.

DISCLOSURE OF THE INVENTION

In the PBD monomer which is not linked to the cell binding agent, the Cring may be prone to become fully aromatic by oxidation(‘aromatization’). This process may be spontaneous, or the aromatizationmay occur during synthesis. For example, sibiromycin:

is known to aromatise, which leads to inactivity of the PBD.Aromatization can be detected using NMR (by determining the present ofan addition aromatic hydrogen).

To overcome this issue, the present inventors have synthesised PBDdimers where the C-ring not linked to the cell binding agent has anexo-double bond, thus retaining the sp² centre at C2.

The present invention comprises a compound with the formula I:

wherein:R² is of formula II′:

where A is a C₅₋₇ aryl group, X is selected from the group consistingof: OH, SH, CO₂H, COH, N═C═O, NHNH₂, CONHNH₂,

and NHR^(N), wherein R^(N) is selected from H and C₁₋₄ alkyl, andeither:(i) Q¹ is a single bond, and Q² is selected from a single bond and—Z—(CH₂)_(n)—, where Z is selected from a single bond, O, S and NH and nis from 1 to 3; or(ii) Q¹ is —CH═CH—, and Q² is a single bond;R¹² is selected from ═CH₂, ═CHR^(D1) and ═CR^(D1)R^(D2), where R^(D1)and R^(D2) are independently selected from R, CO₂R, COR, CHO, CO₂H, andhalo;R⁶ and R⁹ are independently selected from H, R, OH, OR, SH, SR, NH₂,NHR, NRR′, nitro, Me₃Sn and halo;R⁷ is selected from H, R, OH, OR, SH, SR, NH₂, NHR, NHRR′, nitro, Me₃Snand halo; where R and R′ are independently selected from optionallysubstituted C₁₋₁₂ alkyl, C₃₋₂₀ heterocyclyl and C₅₋₂₀ aryl groups;either:(a) R¹⁰ is H, and R¹¹ is OH, OR^(A), where R^(A) is C₁₋₄ alkyl;(b) R¹⁰ and R¹¹ form a nitrogen-carbon double bond between the nitrogenand carbon atoms to which they are bound; or(c) R¹⁰ is H and R¹¹ is SO_(z)M, where z is 2 or 3 and M is a monovalentpharmaceutically acceptable cation;R″ is a C3-12 alkylene group, which chain is optionally interrupted byone or more heteroatoms, e.g. O, S, NR^(N2) (where R^(N2) is H or C₁₋₄alkyl), and/or aromatic rings, e.g. benzene or pyridine;Y and Y′ are selected from 0, S, or NH;R^(6′), R^(7′), R^(9′) are selected from the same groups as R⁶, R⁷ andR⁹ respectively and R^(10′) andR^(11′) are the same as R¹⁰ and R¹¹, wherein if R¹¹ and R^(11′) areSO_(Z)M, M may represent a divalent pharmaceutically acceptable cation.

A second aspect of the present invention provides the use of a compoundof the first aspect of the invention in the manufacture of a medicamentfor treating a proliferative disease. The second aspect also provides acompound of the first aspect of the invention for use in the treatmentof a proliferative disease.

One of ordinary skill in the art is readily able to determine whether ornot a candidate conjugate treats a proliferative condition for anyparticular cell type. For example, assays which may conveniently be usedto assess the activity offered by a particular compound are described inthe examples below.

A third aspect of the present invention comprises a compound of formulaII:

wherein:R² is of formula II′:

where A is a C5-7 aryl group, X is selected from the group comprising:OH, SH, CO₂H, COH, N═C═O, NHNH₂, CONHNH₂,

NHR^(N), wherein R^(N) is selected from the group comprising H and C₁₋₄alkyl, and either:(i) Q¹ is a single bond, and Q² is selected from a single bond and—Z—(CH₂)_(n)—, where Z is selected from a single bond, O, S and NH and nis from 1 to 3; or(ii) Q¹ is —CH═CH—, and Q² is a single bond;R¹² is selected from ═CH₂, ═CHR^(D1) and ═CR^(D1)R^(D2), where R^(D1)and R^(D2) are independently selected from R, CO₂R, COR, CHO, CO₂H, andhalo;R⁶ and R⁹ are independently selected from H, R, OH, OR, SH, SR, NH₂,NHR, NRR′, nitro, Me₃Sn and halo;where R and R′ are independently selected from optionally substitutedC₁₋₁₂ alkyl, C₃₋₂₀ heterocyclyl and C₅₋₂₀ aryl groups;R⁷ is selected from H, R, OH, OR, SH, SR, NH₂, NHR, NHRR′, nitro, Me₃Snand halo;either:(a) R¹⁰ is carbamate nitrogen protecting group, and R¹¹ is O-Prot^(o),wherein Prot^(o) is an oxygen protecting group; or(b) R¹⁰ is a hemi-aminal nitrogen protecting group and R¹¹ is an oxogroup;R″ is a C₃₋₁₂ alkylene group, which chain may be interrupted by one ormore heteroatoms, e.g. O, S, NR^(N2) (where R^(N2) is H or C₁₋₄ alkyl),and/or aromatic rings, e.g. benzene or pyridine;Y and Y′ are selected from 0, S, or NH;R^(6′), R^(7′), R^(9′) are selected from the same groups as R⁶, R⁷ andR⁹ respectively and R^(10′) and R^(11′) are the same as R¹⁰ and R¹¹.

A fourth aspect of the present invention comprises a method of making acompound of formula I from a compound of formula II by deprotection ofthe imine bond.

A fifth aspect of the present invention provides a method of making acompound of the first or third aspect of the invention, comprising atleast one of the method steps set out below.

In a sixth aspect, the present invention relates to Conjugatescomprising dimers of PBDs linked to a targeting agent, wherein a PBD isa dimer of formula I (supra).

In some embodiments, the Conjugates have the following formula III:L-(LU-D)_(p)  (III)wherein L is a Ligand unit (i.e., a targeting agent), LU is a Linkerunit and D is a Drug unit comprising a PBD dimer according to formula I,wherein LU is connected to D via the X substituent of R². The subscriptp is an integer of from 1 to 20. Accordingly, the Conjugates comprise aLigand unit covalently linked to at least one Drug unit by a Linkerunit. The Ligand unit, described more fully below, is a targeting agentthat binds to a target moiety. The Ligand unit can, for example,specifically bind to a cell component (a Cell Binding Agent) or to othertarget molecules of interest. Accordingly, the present invention alsoprovides methods for the treatment of, for example, various cancers andautoimmune disease. These methods encompass the use of the Conjugateswherein the Ligand unit is a targeting agent that specifically binds toa target molecule. The Ligand unit can be, for example, a protein,polypeptide or peptide, such as an antibody, an antigen-binding fragmentof an antibody, or other binding agent, such as an Fc fusion protein.

The PBD dimer D is of formula I, except that X is selected from thegroup comprising: O, S, C(═O), C═, NH(C═O), NHNH, CONHNH,

NR^(N), wherein R^(N) is selected from the group comprising H and C₁₋₄alkyl.

In a seventh aspect of the invention, there is provided a drug linkerhaving formula IV:DLU-D  (IV)wherein DLU is a Drug Linker unit, andD is a Drug unit comprising a PBD dimer according to formula I, whereinDLU is connected to D via the X substituent of R².

BRIEF DESCRIPTION OF FIGURE

FIG. 1 shows the effect of a conjugate of the invention on a tumour.

Definitions

Pharmaceutically acceptable cations Examples of pharmaceuticallyacceptable monovalent and divalent cations are discussed in Berge, etal., J. Pharm. Sci., 66, 1-19 (1977), which is incorporated herein byreference.

The pharmaceutically acceptable cation may be inorganic or organic.

Examples of pharmaceutically acceptable monovalent inorganic cationsinclude, but are not limited to, alkali metal ions such as Na⁺ and K⁺.Examples of pharmaceutically acceptable divalent inorganic cationsinclude, but are not limited to, alkaline earth cations such as Ca²⁺ andMg²⁺. Examples of pharmaceutically acceptable organic cations include,but are not limited to, ammonium ion (i.e. NH₄ ⁺) and substitutedammonium ions (e.g. NH₃R⁺, NH₂R₂ ⁺, NHR₃ ⁺, NR₄ ⁺). Examples of somesuitable substituted ammonium ions are those derived from: ethylamine,diethylamine, dicyclohexylamine, triethylamine, butylamine,ethylenediamine, ethanolamine, diethanolamine, piperazine, benzylamine,phenylbenzylamine, choline, meglumine, and tromethamine, as well asamino acids, such as lysine and arginine. An example of a commonquaternary ammonium ion is N(CH₃)₄ ⁺.

Substituents

The phrase “optionally substituted” as used herein, pertains to a parentgroup which may be unsubstituted or which may be substituted.

Unless otherwise specified, the term “substituted” as used herein,pertains to a parent group which bears one or more substituents. Theterm “substituent” is used herein in the conventional sense and refersto a chemical moiety which is covalently attached to, or if appropriate,fused to, a parent group. A wide variety of substituents are well known,and methods for their formation and introduction into a variety ofparent groups are also well known.

Examples of substituents are described in more detail below.

C₁₋₁₂ alkyl: The term “C₁₋₁₂ alkyl” as used herein, pertains to amonovalent moiety obtained by removing a hydrogen atom from a carbonatom of a hydrocarbon compound having from 1 to 12 carbon atoms, whichmay be aliphatic or alicyclic, and which may be saturated or unsaturated(e.g. partially unsaturated, fully unsaturated). Thus, the term “alkyl”includes the sub-classes alkenyl, alkynyl, cycloalkyl, etc., discussedbelow.

Examples of saturated alkyl groups include, but are not limited to,methyl (C₁), ethyl (C₂), propyl (C₃), butyl (C₄), pentyl (C₅), hexyl(C₆) and heptyl (C₇).

Examples of saturated linear alkyl groups include, but are not limitedto, methyl (C₁), ethyl (C₂), n-propyl (C₃), n-butyl (C4), n-pentyl(amyl) (C₅), n-hexyl (C₆) and n-heptyl (C₇).

Examples of saturated branched alkyl groups include iso-propyl (C₃),iso-butyl (C₄), sec-butyl (C₄), tert-butyl (C₄), iso-pentyl (C₅), andneo-pentyl (C₅).

C₂₋₁₂ Alkenyl: The term “C₂₋₁₂ alkenyl” as used herein, pertains to analkyl group having one or more carbon-carbon double bonds.

Examples of unsaturated alkenyl groups include, but are not limited to,ethenyl (vinyl, —CH═CH₂), 1-propenyl (—CH═CH—CH₃), 2-propenyl (allyl,—CH—CH═CH₂), isopropenyl (1-methylvinyl, —C(CH₃)═CH₂), butenyl (C₄),pentenyl (C₅), and hexenyl (C₆).

C₂₋₁₂ alkynyl: The term “C₂₋₁₂ alkynyl” as used herein, pertains to analkyl group having one or more carbon-carbon triple bonds.

Examples of unsaturated alkynyl groups include, but are not limited to,ethynyl (—C≡CH) and 2-propynyl (propargyl, —CH₂—C≡CH).

C₃₋₁₂ cycloalkyl: The term “C₃₋₁₂ cycloalkyl” as used herein, pertainsto an alkyl group which is also a cyclyl group; that is, a monovalentmoiety obtained by removing a hydrogen atom from an alicyclic ring atomof a cyclic hydrocarbon (carbocyclic) compound, which moiety has from 3to 7 carbon atoms, including from 3 to 7 ring atoms.

Examples of cycloalkyl groups include, but are not limited to, thosederived from:

-   -   saturated monocyclic hydrocarbon compounds:        cyclopropane (C₃), cyclobutane (C₄), cyclopentane (C₅),        cyclohexane (C₆), cycloheptane (C₇), methylcyclopropane (C₄),        dimethylcyclopropane (C₅), methylcyclobutane (C₅),        dimethylcyclobutane (C₆), methylcyclopentane (C₆),        dimethylcyclopentane (C₇) and methylcyclohexane (C₇);    -   unsaturated monocyclic hydrocarbon compounds:        cyclopropene (C₃), cyclobutene (C₄), cyclopentene (C₅),        cyclohexene (C₆), methylcyclopropene (C₄), dimethylcyclopropene        (C₅), methylcyclobutene (C₅), dimethylcyclobutene (C₆),        methylcyclopentene (C₆), dimethylcyclopentene (C₇) and        methylcyclohexene (C₇); and    -   saturated polycyclic hydrocarbon compounds:        norcarane (C₇), norpinane (C₇), norbornane (C₇).        C₃₋₂₀ heterocyclyl: The term “C₃₋₂₀ heterocyclyl” as used        herein, pertains to a monovalent moiety obtained by removing a        hydrogen atom from a ring atom of a heterocyclic compound, which        moiety has from 3 to 20 ring atoms, of which from 1 to 10 are        ring heteroatoms. Preferably, each ring has from 3 to 7 ring        atoms, of which from 1 to 4 are ring heteroatoms.

In this context, the prefixes (e.g. C₃₋₂₀, C₃₋₇, C₅₋₆, etc.) denote thenumber of ring atoms, or range of number of ring atoms, whether carbonatoms or heteroatoms. For example, the term “C₅₋₆heterocyclyl”, as usedherein, pertains to a heterocyclyl group having 5 or 6 ring atoms.

Examples of monocyclic heterocyclyl groups include, but are not limitedto, those derived from:

N₁: aziridine (C₃), azetidine (C₄), pyrrolidine (tetrahydropyrrole)(C₅), pyrroline (e.g., 3-pyrroline, 2,5-dihydropyrrole) (C₅), 2H-pyrroleor 3H-pyrrole (isopyrrole, isoazole) (C₅), piperidine (C₆),dihydropyridine (C₆), tetrahydropyridine (C₆), azepine (C₇);O₁: oxirane (C₃), oxetane (C₄), oxolane (tetrahydrofuran) (C₅), oxole(dihydrofuran) (C₅), oxane (tetrahydropyran) (C₆), dihydropyran (C₆),pyran (C₆), oxepin (C₇);S₁: thiirane (C₃), thietane (C₄), thiolane (tetrahydrothiophene) (C₅),thiane (tetrahydrothiopyran) (C₆), thiepane (C₇);O₂: dioxolane (C₅), dioxane (C₆), and dioxepane (C₇);O₃: trioxane (C₆);N₂: imidazolidine (C₅), pyrazolidine (diazolidine) (C₅), imidazoline(C₅), pyrazoline (dihydropyrazole) (C₅), piperazine (C6);N₁O₁: tetrahydrooxazole (C₅), dihydrooxazole (C₅), tetrahydroisoxazole(C₅), dihydroisoxazole (C₅), morpholine (C₆), tetrahydrooxazine (C₆),dihydrooxazine (C₆), oxazine (C₆);N₁S₁: thiazoline (C₅), thiazolidine (C₅), thiomorpholine (C₆);N₂O₁: oxadiazine (C6);O₁S₁: oxathiole (C₅) and oxathiane (thioxane) (C₆); and,N₁O₁S₁: oxathiazine (C6).

Examples of substituted monocyclic heterocyclyl groups include thosederived from saccharides, in cyclic form, for example, furanoses (C₅),such as arabinofuranose, lyxofuranose, ribofuranose, and xylofuranse,and pyranoses (C₆), such as allopyranose, altropyranose, glucopyranose,mannopyranose, gulopyranose, idopyranose, galactopyranose, andtalopyranose.

C₅₋₂₀ aryl: The term “C₅₋₂₀ aryl”, as used herein, pertains to amonovalent moiety obtained by removing a hydrogen atom from an aromaticring atom of an aromatic compound, which moiety has from 3 to 20 ringatoms. Preferably, each ring has from 5 to 7 ring atoms.

In this context, the prefixes (e.g. C₃₋₂₀, C₅₋₇, C₅₋₆, etc.) denote thenumber of ring atoms, or range of number of ring atoms, whether carbonatoms or heteroatoms. For example, the term “C₅₋₈ aryl” as used herein,pertains to an aryl group having 5 or 6 ring atoms.

The ring atoms may be all carbon atoms, as in “carboaryl groups”.

Examples of carboaryl groups include, but are not limited to, thosederived from benzene (i.e. phenyl) (C₆), naphthalene (C₁₀), azulene(C₁₀), anthracene (C₁₄), phenanthrene (C₁₄), naphthacene (C₁₆), andpyrene (C₁₆).

Examples of aryl groups which comprise fused rings, at least one ofwhich is an aromatic ring, include, but are not limited to, groupsderived from indane (e.g. 2,3-dihydro-1H-indene) (C₉), indene (C₉),isoindene (C₉), tetraline (1,2,3,4-tetrahydronaphthalene (C₁₀),acenaphthene (C₁₂), fluorene (C₁₃), phenalene (C₁₃), acephenanthrene(C₁₅), and aceanthrene (C₁₆).

Alternatively, the ring atoms may include one or more heteroatoms, as in“heteroaryl groups”. Examples of monocyclic heteroaryl groups include,but are not limited to, those derived from:

N₁: pyrrole (azole) (C₅), pyridine (azine) (C₆);

O₁: furan (oxole) (C₅);

S₁: thiophene (thiole) (C₅); N₁O₁: oxazole (C₅), isoxazole (C₅),isoxazine (C₆);

N₂O₁: oxadiazole (furazan) (C₅);

N₃O₁: oxatriazole (C₅);

N₁S₁: thiazole (C₅), isothiazole (C₅);

N₂: imidazole (1,3-diazole) (C₅), pyrazole (1,2-diazole) (C₅),pyridazine (1,2-diazine) (C₆), pyrimidine (1,3-diazine) (C₆) (e.g.,cytosine, thymine, uracil), pyrazine (1,4-diazine) (C₆);

N₃: triazole (C₅), triazine (C₆); and,

N₄: tetrazole (C₅).

Examples of heteroaryl which comprise fused rings, include, but are notlimited to:

-   -   C₉ (with 2 fused rings) derived from benzofuran (O₁),        isobenzofuran (O₁), indole (N₁), isoindole (N₁), indolizine        (N₁), indoline (N₁), isoindoline (N₁), purine (N₄) (e.g.,        adenine, guanine), benzimidazole (N₂), indazole (N₂),        benzoxazole (N₁O₁), benzisoxazole (N₁O₁), benzodioxole (O₂),        benzofurazan (N₂O₁), benzotriazole (N₃), benzothiofuran (S₁),        benzothiazole (N₁S₁), benzothiadiazole (N₂S);    -   C₁₀ (with 2 fused rings) derived from chromene (O₁), isochromene        (O₁), chroman (O₁), isochroman (O₁), benzodioxan (O₂), quinoline        (N₁), isoquinoline (N₁), quinolizine (N₁), benzoxazine (N₁O₁),        benzodiazine (N₂), pyridopyridine (N₂), quinoxaline (N₂),        quinazoline (N₂), cinnoline (N₂), phthalazine (N₂),        naphthyridine (N₂), pteridine (N₄);    -   C₁₁ (with 2 fused rings) derived from benzodiazepine (N₂);    -   C₁₃ (with 3 fused rings) derived from carbazole (N₁),        dibenzofuran (O₁), dibenzothiophene (S₁), carboline (N₂),        perimidine (N₂), pyridoindole (N₂); and,    -   C₁₄ (with 3 fused rings) derived from acridine (N₁), xanthene        (O₁), thioxanthene (S₁), oxanthrene (O₂), phenoxathiin (O₁S₁),        phenazine (N₂), phenoxazine (N₁O₁), phenothiazine (N₁S₁),        thianthrene (S₂), phenanthridine (N₁), phenanthroline (N₂),        phenazine (N₂).

The above groups, whether alone or part of another substituent, maythemselves optionally be substituted with one or more groups selectedfrom themselves and the additional substituents listed below.

Halo: —F, —Cl, —Br, and —I.

Hydroxy: —OH.

Ether: —OR, wherein R is an ether substituent, for example, a C₁₋₇ alkylgroup (also referred to as a C₁₋₇ alkoxy group, discussed below), aC₃₋₂₀ heterocyclyl group (also referred to as a C₃₋₂₀ heterocyclyloxygroup), or a C₃₋₂₀ aryl group (also referred to as a C₃₋₂₃ aryloxygroup), preferably a C₁₋₇alkyl group.

Alkoxy: —OR, wherein R is an alkyl group, for example, a C₁₋₇ alkylgroup. Examples of C₁₋₇ alkoxy groups include, but are not limited to,—OMe (methoxy), —OEt (ethoxy), —O(nPr) (n-propoxy), —O(iPr)(isopropoxy), —O(nBu) (n-butoxy), —O(sBu) (sec-butoxy), —O(iBu)(isobutoxy), and —O(tBu) (tert-butoxy).

Acetal: —CH(OR¹)(OR²), wherein R¹ and R² are independently acetalsubstituents, for example, a C₁₋₇ alkyl group, a C₃₋₂₀ heterocyclylgroup, or a C₅₋₂₀ aryl group, preferably a C₁₋₇ alkyl group, or, in thecase of a “cyclic” acetal group, R¹ and R², taken together with the twooxygen atoms to which they are attached, and the carbon atoms to whichthey are attached, form a heterocyclic ring having from 4 to 8 ringatoms. Examples of acetal groups include, but are not limited to,—CH(OMe)₂, —CH(OEt)₂, and —CH(OMe)(OEt).

Hemiacetal: —CH(OH)(OR¹), wherein R¹ is a hemiacetal substituent, forexample, a C₁₋₇ alkyl group, a C₃₋₂₀ heterocyclyl group, or a C₅₋₂₀ arylgroup, preferably a C₁₋₇ alkyl group. Examples of hemiacetal groupsinclude, but are not limited to, —CH(OH)(OMe) and —CH(OH)(OEt).

Ketal: —CR(OR¹)(OR²), where R¹ and R² are as defined for acetals, and Ris a ketal substituent other than hydrogen, for example, a C₁₋₇ alkylgroup, a C₃₋₂₀ heterocyclyl group, or a C₅₋₂₀ aryl group, preferably aC₁₋₇ alkyl group. Examples ketal groups include, but are not limited to,—C(Me)(OMe)₂, —C(Me)(OEt)₂, —C(Me)(OMe)(OEt), —C(Et)(OMe)₂,—C(Et)(OEt)₂, and —C(Et)(OMe)(OEt).

Hemiketal: —CR(OH)(OR¹), where R¹ is as defined for hemiacetals, and Ris a hemiketal substituent other than hydrogen, for example, a C₁₋₇alkyl group, a C₃₋₂₀ heterocyclyl group, or a C₅₋₂₀ aryl group,preferably a C₁₋₇ alkyl group. Examples of hemiacetal groups include,but are not limited to, —C(Me)(OH)(OMe), —C(Et)(OH)(OMe),—C(Me)(OH)(OEt), and —C(Et)(OH)(OEt).

Oxo (keto, -one): ═O.

Thione (thioketone): ═S.

Imino (imine): ═NR, wherein R is an imino substituent, for example,hydrogen, C₁₋₇ alkyl group, a C₃₋₂₀ heterocyclyl group, or a C₅₋₂₀ arylgroup, preferably hydrogen or a C₁₋₇ alkyl group. Examples of estergroups include, but are not limited to, ═NH, ═NMe, ═NEt, and ═NPh.

Formyl (carbaldehyde, carboxaldehyde): —C(═O)H.

Acyl (keto): —C(═O)R, wherein R is an acyl substituent, for example, aC₁₋₇ alkyl group (also referred to as C₁₋₇ alkylacyl or C₁₋₇ alkanoyl),a C₃₋₂₀ heterocyclyl group (also referred to as C₃₋₂₀ heterocyclylacyl),or a C₅₋₂₀ aryl group (also referred to as C₅₋₂₀ arylacyl), preferably aC₁₋₇ alkyl group. Examples of acyl groups include, but are not limitedto, —C(═O)CH₃ (acetyl), —C(═O)CH₂CH₃ (propionyl), —C(═O)C(CH₃)₃(t-butyryl), and —C(═O)Ph (benzoyl, phenone).

Carboxy (carboxylic acid): —C(═O)OH.

Thiocarboxy (thiocarboxylic acid): —C(═S)SH.

Thiolocarboxy (thiolocarboxylic acid): —C(═O)SH.

Thionocarboxy (thionocarboxylic acid): —C(═S)OH.

Imidic acid: —C(═NH)OH.

Hydroxamic acid: —C(═NOH)OH.

Ester (carboxylate, carboxylic acid ester, oxycarbonyl): —C(═O)OR,wherein R is an ester substituent, for example, a C₁₋₇ alkyl group, aC₃₋₂₀ heterocyclyl group, or a C₅₋₂₀ aryl group, preferably a C₁₋₇ alkylgroup. Examples of ester groups include, but are not limited to,—C(═O)OCH₃, —C(═O)OCH₂CH₃, —C(═O)OC(CH₃)₃, and —C(═O)OPh.

Acyloxy (reverse ester): —OC(═O)R, wherein R is an acyloxy substituent,for example, a C₁₋₇ alkyl group, a C₃₋₂₀ heterocyclyl group, or a C₅₋₂₀aryl group, preferably a C₁₋₇ alkyl group. Examples of acyloxy groupsinclude, but are not limited to, —OC(═O)CH₃ (acetoxy), —OC(═O)CH₂CH₃,—OC(═O)C(CH₃)₃, —OC(═O)Ph, and —OC(═O)CH₂Ph.

Oxycarboyloxy: —OC(═O)OR, wherein R is an ester substituent, forexample, a C₁₋₇ alkyl group, a C₃₋₂₀ heterocyclyl group, or a C₅₋₂₀ arylgroup, preferably a C₁₋₇ alkyl group. Examples of ester groups include,but are not limited to, —OC(═O)OCH₃, —OC(═O)OCH₂CH₃, —OC(═O)OC(CH₃)₃,and —OC(═O)OPh.

Amino: —NR¹R², wherein R¹ and R² are independently amino substituents,for example, hydrogen, a C₁₋₇ alkyl group (also referred to as C₁₋₇alkylamino or di-C₁₋₇ alkylamino), a C₃₋₂₀ heterocyclyl group, or aC₅₋₂₀ aryl group, preferably H or a C₁₋₇ alkyl group, or, in the case ofa “cyclic” amino group, R¹ and R², taken together with the nitrogen atomto which they are attached, form a heterocyclic ring having from 4 to 8ring atoms. Amino groups may be primary (—NH₂), secondary (—NHR¹), ortertiary (—NHR¹R²), and in cationic form, may be quaternary (—⁺NR¹R²R³).Examples of amino groups include, but are not limited to, —NH₂, —NHCH₃,—NHC(CH₃)₂, —N(CH₃)₂, —N(CH₂CH₃)₂, and —NHPh. Examples of cyclic aminogroups include, but are not limited to, aziridino, azetidino,pyrrolidino, piperidino, piperazino, morpholino, and thiomorpholino.

Amido (carbamoyl, carbamyl, aminocarbonyl, carboxamide): —C(═O)NR¹R²,wherein R¹ and R² are independently amino substituents, as defined foramino groups. Examples of amido groups include, but are not limited to,—C(═O)NH₂, —C(═O)NHCH₃, —C(═O)N(CH₃)₂, —C(═O)NHCH₂CH₃, and—C(═O)N(CH₂CH₃)₂, as well as amido groups in which R¹ and R², togetherwith the nitrogen atom to which they are attached, form a heterocyclicstructure as in, for example, piperidinocarbonyl, morpholinocarbonyl,thiomorpholinocarbonyl, and piperazinocarbonyl.

Thioamido (thiocarbamyl): —C(═S)NR¹R², wherein R¹ and R² areindependently amino substituents, as defined for amino groups. Examplesof amido groups include, but are not limited to, —C(═S)NH₂, —C(═S)NHCH₃,—C(═S)N(CH₃)₂, and —C(═S)NHCH₂CH₃.

Acylamido (acylamino): —NR′C(═O)R², wherein R′ is an amide substituent,for example, hydrogen, a C₁₋₇ alkyl group, a C₃₋₂₀ heterocyclyl group,or a C₅₋₂₀ aryl group, preferably hydrogen or a C₁₋₇ alkyl group, and R²is an acyl substituent, for example, a C₁₋₇ alkyl group, a C₃₋₂₀heterocyclyl group, or a C₅₋₂₀aryl group, preferably hydrogen or a C₁₋₇alkyl group. Examples of acylamide groups include, but are not limitedto, —NHC(═O)CH₃, —NHC(═O)CH₂CH₃, and —NHC(═O)Ph. R¹ and R² may togetherform a cyclic structure, as in, for example, succinimidyl, maleimidyl,and phthalimidyl:

Aminocarbonyloxy: —OC(═O)NR¹R², wherein R¹ and R² are independentlyamino substituents, as defined for amino groups. Examples ofaminocarbonyloxy groups include, but are not limited to, —OC(═O)NH₂,—OC(═O)NHMe, —OC(═O)NMe₂, and —OC(═O)NEt₂.

Ureido: —N(R¹)CONR²R³ wherein R² and R³ are independently aminosubstituents, as defined for amino groups, and R′ is a ureidosubstituent, for example, hydrogen, a C₁₋₇ alkyl group, aC₃₋₂₀heterocyclyl group, or a C₅₋₂₀ aryl group, preferably hydrogen or aC₁₋₇ alkyl group. Examples of ureido groups include, but are not limitedto, —NHCONH₂, —NHCONHMe, —NHCONHEt, —NHCONMe₂, —NHCONEt₂, —NMeCONH₂,—NMeCONHMe, —NMeCONHEt, —NMeCONMe₂, and —NMeCONEt₂.

Guanidino: —NH—C(═NH)NH₂.

Tetrazolyl: a five membered aromatic ring having four nitrogen atoms andone carbon atom,

Imino: =NR, wherein R is an imino substituent, for example, for example,hydrogen, a C₁₋₇ alkyl group, a C₃₋₂₀heterocyclyl group, or a C₅₋₂₀ arylgroup, preferably H or a C₁₋₇ alkyl group. Examples of imino groupsinclude, but are not limited to, ═NH, ═NMe, and ═NEt.

Amidine (amidino): —C(═NR)NR₂, wherein each R is an amidine substituent,for example, hydrogen, a C₁₋₇ alkyl group, a C₃₋₂₀heterocyclyl group, ora C₅₋₂₀ aryl group, preferably H or a C1-7 alkyl group. Examples ofamidine groups include, but are not limited to, —C(═NH)NH₂, —C(═NH)NMe₂,and —C(═NMe)NMe₂.

Nitro: —NO₂.

Nitroso: —NO.

Azido: —N₃.

Cyano (nitrile, carbonitrile): —CN.

Isocyano: —NC.

Cyanato: —OCN.

Isocyanato: —NCO.

Thiocyano (thiocyanato): —SCN.

Isothiocyano (isothiocyanato): —NCS.

Sulfhydryl (thiol, mercapto): —SH.

Thioether (sulfide): —SR, wherein R is a thioether substituent, forexample, a C₁₋₇ alkyl group (also referred to as a C₁₋₇alkylthio group),a C₃₋₂₀ heterocyclyl group, or a C₅₋₂₀ aryl group, preferably a C₁₋₇alkyl group. Examples of C₁₋₇ alkylthio groups include, but are notlimited to, —SCH₃ and —SCH₂CH₃.

Disulfide: —SS—R, wherein R is a disulfide substituent, for example, aC₁₋₇ alkyl group, a C₃₋₂₀ heterocyclyl group, or a C₅₋₂₀ aryl group,preferably a C₁₋₇ alkyl group (also referred to herein as C₁₋₇ alkyldisulfide). Examples of C₁₋₇ alkyl disulfide groups include, but are notlimited to, —SSCH₃ and —SSCH₂CH₃.

Sulfine (sulfinyl, sulfoxide): —S(═O)R, wherein R is a sulfinesubstituent, for example, a C₁₋₇ alkyl group, a C₃₋₂₀ heterocyclylgroup, or a C₅₋₂₀ aryl group, preferably a C₁₋₇ alkyl group. Examples ofsulfine groups include, but are not limited to, —S(═O)CH₃ and—S(═O)CH₂CH₃.

Sulfone (sulfonyl): —S(═O)₂R, wherein R is a sulfone substituent, forexample, a C₁₋₇ alkyl group, a C₃₋₂₀ heterocyclyl group, or a C₅₋₂₀ arylgroup, preferably a C₁₋₇ alkyl group, including, for example, afluorinated or perfluorinated C₁₋₇ alkyl group. Examples of sulfonegroups include, but are not limited to, —S(═O)₂CH₃ (methanesulfonyl,mesyl), —S(═O)₂CF₃ (triflyl), —S(═O)₂CH₂CH₃ (esyl), —S(═O)₂C4F₉(nonaflyl), —S(═O)₂CH₂CF₃ (tresyl), —S(═O)₂CH₂CH₂NH₂ (tauryl), —S(═O)₂Ph(phenylsulfonyl, besyl), 4-methylphenylsulfonyl (tosyl),4-chlorophenylsulfonyl (closyl), 4-bromophenylsulfonyl (brosyl),4-nitrophenyl (nosyl), 2-naphthalenesulfonate (napsyl), and5-dimethylamino-naphthalen-1-ylsulfonate (dansyl).

Sulfinic acid (sulfino): —S(═O)OH, —SO₂H.

Sulfonic acid (sulfo): —S(═O)20H, —SO₃H.

Sulfinate (sulfinic acid ester): —S(═O)OR; wherein R is a sulfinatesubstituent, for example, a C₁₋₇ alkyl group, a C₃₋₂₀ heterocyclylgroup, or a C₅₋₂₀ aryl group, preferably a C₁₋₇ alkyl group. Examples ofsulfinate groups include, but are not limited to, —S(═O)OCH₃(methoxysulfinyl; methyl sulfinate) and —S(═O)OCH₂CH₃ (ethoxysulfinyl;ethyl sulfinate).

Sulfonate (sulfonic acid ester): —S(═O)₂OR, wherein R is a sulfonatesubstituent, for example, a C₁₋₇ alkyl group, a C₃₋₂₀ heterocyclylgroup, or a C₅₋₂₀ aryl group, preferably a C₁₋₇ alkyl group. Examples ofsulfonate groups include, but are not limited to, —S(═O)₂OCH₃(methoxysulfonyl; methyl sulfonate) and —S(═O)₂OCH₂CH₃ (ethoxysulfonyl;ethyl sulfonate).

Sulfinyloxy: —OS(═O)R, wherein R is a sulfinyloxy substituent, forexample, a C₁₋₇ alkyl group, a C₃₋₂₀ heterocyclyl group, or a C₅₋₂₀ arylgroup, preferably a C₁₋₇ alkyl group.

Examples of sulfinyloxy groups include, but are not limited to,—OS(═O)CH₃ and —OS(═O)CH₂CH₃.

Sulfonyloxy: —OS(═O)2R, wherein R is a sulfonyloxy substituent, forexample, a C₁₋₇ alkyl group, a C₃₋₂₀ heterocyclyl group, or a C₅₋₂₀ arylgroup, preferably a C₁₋₇ alkyl group. Examples of sulfonyloxy groupsinclude, but are not limited to, —OS(═O)₂CH₃ (mesylate) and—OS(═O)₂CH₂CH₃ (esylate).

Sulfate: —OS(═O)₂OR; wherein R is a sulfate substituent, for example, aC₁₋₇ alkyl group, a C₃₋₂₀ heterocyclyl group, or a C₅₋₂₀ aryl group,preferably a C₁₋₇ alkyl group. Examples of sulfate groups include, butare not limited to, —OS(═O)₂OCH₃ and —SO(═O)₂OCH₂CH₃.

Sulfamyl (sulfamoyl; sulfinic acid amide; sulfinamide): —S(═O)NR¹R²,wherein R¹ and R² are independently amino substituents, as defined foramino groups. Examples of sulfamyl groups include, but are not limitedto, —S(═O)NH₂, —S(═O)NH(CH₃), —S(═O)N(CH₃)₂, —S(═O)NH(CH₂CH₃),—S(═O)N(CH₂CH₃)₂, and —S(═O)NHPh.

Sulfonamido (sulfinamoyl; sulfonic acid amide; sulfonamide):—S(═O)₂NR¹R², wherein R¹ and R² are independently amino substituents, asdefined for amino groups. Examples of sulfonamido groups include, butare not limited to, —S(═O)₂NH₂, —S(═O)₂NH(CH₃), —S(═O)₂N(CH₃)₂,—S(═O)₂NH(CH₂CH₃), —S(═O)₂N(CH₂CH₃)₂, and —S(═O)₂NHPh.

Sulfamino: —NR¹S(═O)₂OH, wherein R′ is an amino substituent, as definedfor amino groups. Examples of sulfamino groups include, but are notlimited to, —NHS(═O)₂OH and —N(CH₃)S(═O)₂OH.

Sulfonamino: —NR¹S(═O)₂R, wherein R¹ is an amino substituent, as definedfor amino groups, and R is a sulfonamino substituent, for example, aC₁₋₇ alkyl group, a C₃₋₂₀ heterocyclyl group, or a C₅₋₂₀ aryl group,preferably a C₁₋₇ alkyl group. Examples of sulfonamino groups include,but are not limited to, —NHS(═O)₂CH₃ and —N(CH₃)S(═O)₂C₆H₅.

Sulfinamino: —NR¹S(═O)R, wherein R¹ is an amino substituent, as definedfor amino groups, and R is a sulfinamino substituent, for example, aC₁₋₇ alkyl group, a C₃₋₂₀ heterocyclyl group, or a C₅₋₂₀ aryl group,preferably a C₁₋₇ alkyl group. Examples of sulfinamino groups include,but are not limited to, —NHS(═O)CH₃ and —N(CH₃)S(═O)C₆H₅.

Phosphino (phosphine): —PR₂, wherein R is a phosphino substituent, forexample, —H, a C₁₋₇ alkyl group, a C₃₋₂₀ heterocyclyl group, or a C₅₋₂₀aryl group, preferably —H, a C₁₋₇ alkyl group, or a C₅₋₂₀ aryl group.Examples of phosphino groups include, but are not limited to, —PH₂,—P(CH₃)₂, —P(CH₂CH₃)₂, —P(t-Bu)₂, and —P(Ph)₂.

Phospho: —P(═O)₂.

Phosphinyl (phosphine oxide): —P(═O)R₂, wherein R is a phosphinylsubstituent, for example, a C₁₋₇ alkyl group, a C₃₋₂₀ heterocyclylgroup, or a C₅₋₂₀ aryl group, preferably a C₁₋₇ alkyl group or a C₅₋₂₀aryl group. Examples of phosphinyl groups include, but are not limitedto, —P(═O)(CH₃)₂, —P(═O)(CH₂CH₃)₂, —P(═O)(t-Bu)₂, and —P(═O)(Ph)₂.

Phosphonic acid (phosphono): —P(═O)(OH)₂.

Phosphonate (phosphono ester): —P(═O)(OR)₂, where R is a phosphonatesubstituent, for example, —H, a C₁₋₇ alkyl group, a C₃₋₂₀ heterocyclylgroup, or a C₅₋₂₀ aryl group, preferably —H, a C₁₋₇ alkyl group, or aC₅₋₂₀ aryl group. Examples of phosphonate groups include, but are notlimited to, —P(═O)(OCH₃)₂, —P(═O)(OCH₂CH₃)₂, —P(═O)(O-t-Bu)₂, and—P(═O)(OPh)₂.

Phosphoric acid (phosphonooxy): —OP(═O)(OH)₂.

Phosphate (phosphonooxy ester): —OP(═O)(OR)₂, where R is a phosphatesubstituent, for example, —H, a C₁₋₇ alkyl group, a C₃₋₂₀ heterocyclylgroup, or a C₃₋₂₀ aryl group, preferably —H, a C₁₋₇ alkyl group, or aC₅₋₂₀ aryl group. Examples of phosphate groups include, but are notlimited to, —OP(═O)(OCH₃)₂, —OP(═O)(OCH₂CH₃)₂, —OP(═O)(O-t-Bu)₂, and—OP(═O)(OPh)₂.

Phosphorous acid: —OP(OH)₂.

Phosphite: —OP(OR)₂, where R is a phosphite substituent, for example,—H, a C₁₋₇ alkyl group, a C₃₋₂₀ heterocyclyl group, or a C₃₋₂₀ arylgroup, preferably —H, a C₁₋₇ alkyl group, or a C₅₋₂₀ aryl group.Examples of phosphite groups include, but are not limited to,—OP(OCH₃)₂, —OP(OCH₂CH₃)₂, —OP(O-t-Bu)₂, and —OP(OPh)₂.

Phosphoramidite: —OP(OR¹)—NR² ₂, where R¹ and R² are phosphoramiditesubstituents, for example, —H, a (optionally substituted) C₁₋₇ alkylgroup, a C₃₋₂₀ heterocyclyl group, or a C₅₋₂₀ aryl group, preferably —H,a C₁₋₇ alkyl group, or a C₅₋₂₀ aryl group. Examples of phosphoramiditegroups include, but are not limited to, —OP(OCH₂CH₃)—N(CH₃)₂,—OP(OCH₂CH₃)—N(i-Pr)₂, and —OP(OCH₂CH₂CN)—N(i-Pr)₂.

Phosphoramidate: —OP(═O)(OR¹)—NR² ₂, where R¹ and R² are phosphoramidatesubstituents, for example, —H, a (optionally substituted) C₁₋₇ alkylgroup, a C₃₋₂₀ heterocyclyl group, or a C₅₋₂₀ aryl group, preferably —H,a C₁₋₇ alkyl group, or a C₅₋₂₀ aryl group. Examples of phosphoramidategroups include, but are not limited to, —OP(═O)(OCH₂CH₃)—N(CH₃)₂,—OP(═O)(OCH₂CH₃)—N(i-Pr)₂, and —OP(═O)(OCH₂CH₂CN)—N(i-Pr)₂.

Alkylene

C₃₋₁₂ alkylene: The term “C₃₋₁₂ alkylene”, as used herein, pertains to abidentate moiety obtained by removing two hydrogen atoms, either bothfrom the same carbon atom, or one from each of two different carbonatoms, of a hydrocarbon compound having from 3 to 12 carbon atoms(unless otherwise specified), which may be aliphatic or alicyclic, andwhich may be saturated, partially unsaturated, or fully unsaturated.Thus, the term “alkylene” includes the sub-classes alkenylene,alkynylene, cycloalkylene, etc., discussed below.

Examples of linear saturated C₃₋₁₂ alkylene groups include, but are notlimited to, —(CH₂)_(n)—where n is an integer from 3 to 12, for example,—CH₂CH₂CH₂— (propylene), —CH₂CH₂CH₂CH₂— (butylene), —CH₂CH₂CH₂CH₂CH₂—(pentylene) and —CH₂CH₂CH₂CH—₂CH₂CH₂CH₂— (heptylene).

Examples of branched saturated C₃₋₁₂ alkylene groups include, but arenot limited to, —CH(CH₃)CH₂—, —CH(CH₃)CH₂CH₂—, —CH(CH₃)CH₂CH₂CH₂—,—CH₂CH(CH₃)CH₂—, —CH₂CH(CH₃)CH₂CH₂—, —CH(CH₂CH₃)—, —CH(CH₂CH₃)CH₂—, and—CH₂CH(CH₂CH₃)CH₂—.

Examples of linear partially unsaturated C₃₋₁₂ alkylene groups (C₃₋₁₂alkenylene, and alkynylene groups) include, but are not limited to,—CH═CH—CH₂—, —CH₂—CH═CH₂—, —CH═CH—CH₂—CH₂—, —CH═CH—CH₂—CH₂—CH₂—,—CH═CH—CH═CH—, —CH═CH—CH═CH—CH₂—, —CH═CH—CH═CH—CH₂—CH₂—,—CH═CH—CH₂—CH═CH—, —CH═CH—CH₂—CH₂—CH═CH—, and —CH₂—C≡C—CH₂—.

Examples of branched partially unsaturated C₃₋₁₂ alkylene groups (C₃₋₁₂alkenylene and alkynylene groups) include, but are not limited to,—C(CH₃)═CH—, —C(CH₃)═CH—CH₂—, —CH═CH—CH(CH₃)— and —C≡C—CH(CH₃)—.

Examples of alicyclic saturated C₃₋₁₂ alkylene groups (C₃₋₁₂cycloalkylenes) include, but are not limited to, cyclopentylene (e.g.cyclopent-1,3-ylene), and cyclohexylene (e.g. cyclohex-1,4-ylene).

Examples of alicyclic partially unsaturated C₃₋₁₂ alkylene groups (C₃₋₁₂cycloalkylenes) include, but are not limited to, cyclopentenylene (e.g.4-cyclopenten-1,3-ylene), cyclohexenylene (e.g. 2-cyclohexen-1,4-ylene;3-cyclohexen-1,2-ylene; 2,5-cyclohexadien-1,4-ylene).

Oxygen protecting group: the term “oxygen protecting group” refers to amoiety which masks a hydroxy group, and these are well known in the art.A large number of suitable groups are described on pages 23 to 200 ofGreene, T. W. and Wuts, G.M., Protective Groups in Organic Synthesis,3^(rd) Edition, John Wiley & Sons, Inc., 1999, which is incorporatedherein by reference. Classes of particular interest include silyl ethers(e.g. TMS, TBDMS), substituted methyl ethers (e.g. THP) and esters (e.g.acetate).

Carbamate nitrogen protecting group: the term “carbamate nitrogenprotecting group” pertains to a moiety which masks the nitrogen in theimine bond, and these are well known in the art. These groups have thefollowing structure:

wherein R′¹⁰ is R as defined above. A large number of suitable groupsare described on pages 503 to 549 of Greene, T. W. and Wuts, G.M.,Protective Groups in Organic Synthesis, 3^(rd) Edition, John Wiley &Sons, Inc., 1999, which is incorporated herein by reference.

Hemi-aminal nitrogen protecting group: the term “hemi-aminal nitrogenprotecting group” pertains to a group having the following structure:

wherein R′¹⁰ is R as defined above. A large number of suitable groupsare described on pages 633 to 647 as amide protecting groups of Greene,T. W. and Wuts, G.M., Protective Groups in Organic Synthesis, 3^(rd)Edition, John Wiley & Sons, Inc., 1999, which is incorporated herein byreference.

Conjugates

The present invention provides Conjugates comprising a PBD dimerconnected to a Ligand unit via a Linker Unit. In one embodiment, theLinker unit includes a Stretcher unit (A), a Specificity unit (L¹), anda Spacer unit (L²). The Linker unit is connected at one end to theLigand unit and at the other end to the PBD dimer compound.

In one aspect, such a Conjugate is shown below in formula IIIa:L-(A¹ _(a)-L¹ _(s)-L² _(y)D)_(p)  (IIIa)

-   -   wherein:    -   L is the Ligand unit; and    -   -A¹ _(a)-L¹ _(s)-L² _(y)- is a Linker unit (LU), wherein:    -   -A¹- is a Stretcher unit,    -   a is 1 or 2,    -   L¹- is a Specificity unit,    -   s is an integer ranging from 1 to 12,    -   -L²- is a Spacer unit,    -   y is 0, 1 or 2;    -   -D is an PBD dimer; and    -   p is from 1 to 20.

In some embodiments of this aspect, a is 1, s is 1 and y is 0, such thatthe conjugate is of formula IIIa-1:L-(A¹-L¹-D)_(p)  (IIIa-1)

In another aspect, such a Conjugate is shown below in formula IIIb:

Also illustrated as:L-(A¹ _(a)-L² _(y)(-L¹ _(s))-D)_(p)  (IIIb)

-   -   wherein:    -   L is the Ligand unit; and    -   -A¹ _(a)-L¹ _(s)(L² _(y))- is a Linker unit (LU), wherein:    -   -A¹- is a Stretcher unit linked to a Spacer unit (L²),    -   a is 1 or 2,    -   L¹- is a Specificity unit linked to a Spacer unit (L²),    -   s is an integer ranging from 0 to 12,    -   -L²- is a Spacer unit,    -   y is 0, 1 or 2;    -   -D is a PBD dimer; and    -   p is from 1 to 20.

Preferences

The following preferences may apply to all aspects of the invention asdescribed above, or may relate to a single aspect. The preferences maybe combined together in any combination.

In one embodiment, the Conjugate has the formula:L-(A¹ _(a)-L¹ _(s)-L² _(y)D)_(p)

-   -   wherein L, A¹, a, L¹, s, L², D and p are as described above.

In one embodiment, the Ligand unit (L) is a Cell Binding Agent (CBA)that specifically binds to a target molecule on the surface of a targetcell.

An exemplary formula for the conjugate is illustrated below:

-   -   where the asterisk indicates the point of attachment to the Drug        unit (D), CBA is the Cell Binding Agent, L¹ is a Specificity        unit, A¹ is a Stretcher unit connecting L¹ to the Cell Binding        Agent, L² is a Spacer unit, which is a covalent bond, a        self-immolative group or together with —OC(═O)— forms a        self-immolative group, and L² is optional.

Another exemplary formula is illustrated below:CBA-A¹ _(a)-L¹ _(s)-L² _(y)-*

-   -   where the asterisk indicates the point of attachment to the Drug        unit (D), CBA is the Cell Binding Agent, L¹ is a Specificity        unit, A¹ is a Stretcher unit connecting L¹ to the Cell Binding        Agent, L² is a Spacer unit which is a covalent bond or a        self-immolative group, and a is 1 or 2, s is 0, 1 or 2, and y is        0 or 1 or 2.

A further exemplary formula is illustrated below:CBA-A¹-L¹-*where the asterisk indicates the point of attachment to the Drug unit(D), CBA is the Cell Binding Agent, L¹ is a Specificity unit and A¹ is aStretcher unit connecting L¹ to the Cell Binding Agent.

In the embodiments illustrated above, L¹ can be a cleavable Specificityunit, and may be referred to as a “trigger” that when cleaved activatesa self-immolative group (or self-immolative groups) L², when aself-immolative group(s) is present. When the Specificity unit L′ iscleaved, or the linkage (i.e., the covalent bond) between L¹ and L² iscleaved, the self-immolative group releases the Drug unit (D).

In another embodiment, the Ligand unit (L) is a Cell Binding Agent (CBA)that specifically binds to a target molecule on the surface of a targetcell. An exemplary formula is illustrated below:

where the asterisk indicates the point of attachment to the Drug (D),CBA is the Cell Binding Agent, L¹ is a Specificity unit connected to L²,A¹ is a Stretcher unit connecting L² to the Cell Binding Agent, L² is aself-immolative group, and a is 1 or 2, s is 1 or 2, and y is 1 or 2.

In the various embodiments discussed herein, the nature of L¹ and L² canvary widely. These groups are chosen on the basis of theircharacteristics, which may be dictated in part, by the conditions at thesite to which the conjugate is delivered. Where the Specificity unit L¹is cleavable, the structure and/or sequence of L¹ is selected such thatit is cleaved by the action of enzymes present at the target site (e.g.,the target cell). L¹ units that are cleavable by changes in pH (e.g.acid or base labile), temperature or upon irradiation (e.g. photolabile)may also be used. L¹ units that are cleavable under reducing oroxidising conditions may also find use in the Conjugates.

In some embodiments, L¹ may comprise one amino acid or a contiguoussequence of amino acids. The amino acid sequence may be the targetsubstrate for an enzyme.

In one embodiment, L¹ is cleavable by the action of an enzyme. In oneembodiment, the enzyme is an esterase or a peptidase. For example, L¹may be cleaved by a lysosomal protease, such as a cathepsin.

In one embodiment, L² is present and together with —C(═O)O— forms aself-immolative group or self-immolative groups. In some embodiments,—C(═O)O— also is a self-immolative group.

In one embodiment, where L¹ is cleavable by the action of an enzyme andL² is present, the enzyme cleaves the bond between L¹ and L², wherebythe self-immolative group(s) release the Drug unit.

L′ and L², where present, may be connected by a bond selected from:

-   -   —C(═O)NH—,    -   —C(═O)O—,    -   —NHC(═O)—,    -   —OC(═O)—,    -   —OC(═O)O—,    -   —NHC(═O)O—,    -   —OC(═O)NH—,    -   —NHC(═O)NH, and    -   —O— (a glycosidic bond).

An amino group of L¹ that connects to L² may be the N-terminus of anamino acid or may be derived from an amino group of an amino acid sidechain, for example a lysine amino acid side chain.

A carboxyl group of L¹ that connects to L² may be the C-terminus of anamino acid or may be derived from a carboxyl group of an amino acid sidechain, for example a glutamic acid amino acid side chain.

A hydroxy group of L¹ that connects to L² may be derived from a hydroxygroup of an amino acid side chain, for example a serine amino acid sidechain.

In one embodiment, —C(═O)O— and L² together form the group:

-   -   where the asterisk indicates the point of attachment to the Drug        unit, the wavy line indicates the point of attachment to the L¹,        Y is —N(H)—, —O—, —C(═O)N(H)— or —C(═O)O—, and n is 0 to 3. The        phenylene ring is optionally substituted with one, two or three        substituents as described herein.

In one embodiment, Y is NH.

In one embodiment, n is 0 or 1. Preferably, n is 0.

Where Y is NH and n is 0, the self-immolative group may be referred toas a p-aminobenzylcarbonyl linker (PABC).

The self-immolative group will allow for release of the Drug unit (i.e.,the asymmetric PBD) when a remote site in the linker is activated,proceeding along the lines shown below (for n=0):

-   -   where the asterisk indicates the attachment to the Drug, L* is        the activated form of the remaining portion of the linker and        the released Drug unit is not shown. These groups have the        advantage of separating the site of activation from the Drug.

In another embodiment, —C(═O)O— and L² together form a group selectedfrom:

-   -   where the asterisk, the wavy line, Y, and n are as defined        above. Each phenylene ring is optionally substituted with one,        two or three substituents as described herein. In one        embodiment, the phenylene ring having the Y substituent is        optionally substituted and the phenylene ring not having the Y        substituent is unsubstituted.

In another embodiment, —C(═O)O— and L² together form a group selectedfrom:

-   -   where the asterisk, the wavy line, Y, and n are as defined        above, E is O, S or NR, D is N, CH, or CR, and F is N, CH, or        CR.

In one embodiment, D is N.

In one embodiment, D is CH.

In one embodiment, E is O or S.

In one embodiment, F is CH.

In a preferred embodiment, the covalent bond between L¹ and L² is acathepsin labile (e.g., cleavable) bond.

In one embodiment, L¹ comprises a dipeptide. The amino acids in thedipeptide may be any combination of natural amino acids and non-naturalamino acids. In some embodiments, the dipeptide comprises natural aminoacids. Where the linker is a cathepsin labile linker, the dipeptide isthe site of action for cathepsin-mediated cleavage. The dipeptide thenis a recognition site for cathepsin.

In one embodiment, the group —X₁—X₂— in dipeptide, —NH—X₁—X₂—CO—, isselected from:

-   -   -Phe-Lys-,    -   Val-Ala-,    -   Val-Lys-,    -   Ala-Lys-,    -   Val-Cit-,    -   Phe-Cit-,    -   Leu-Cit-,    -   Ile-Cit-,    -   Phe-Arg-, and    -   Trp-Cit-;        where Cit is citrulline. In such a dipeptide, —NH— is the amino        group of X₁, and CO is the carbonyl group of X₂.

Preferably, the group —X₁—X₂— in dipeptide, —NH—X₁—X₂—CO—, is selectedfrom:

-   -   -Phe-Lys-,    -   Val-Ala-,    -   Val-Lys-,    -   Ala-Lys-, and    -   Val-Cit-.

Most preferably, the group —X₁—X₂— in dipeptide, —NH—X₁-X₂—CO—, is-Phe-Lys-, Val-Cit or -Val-Ala-.

Other dipeptide combinations of interest include:

-   -   -Gly-Gly-,    -   Pro-Pro-, and    -   Val-Glu-.

Other dipeptide combinations may be used, including those described byDubowchik et al., which is incorporated herein by reference.

In one embodiment, the amino acid side chain is chemically protected,where appropriate. The side chain protecting group may be a group asdiscussed below. Protected amino acid sequences are cleavable byenzymes. For example, a dipeptide sequence comprising a Boc sidechain-protected Lys residue is cleavable by cathepsin.

Protecting groups for the side chains of amino acids are well known inthe art and are described in the Novabiochem Catalog. Additionalprotecting group strategies are set out in Protective groups in OrganicSynthesis, Greene and Wuts.

Possible side chain protecting groups are shown below for those aminoacids having reactive side chain functionality:

-   -   Arg: Z, Mtr, Tos;    -   Asn: Trt, Xan;    -   Asp: Bzl, t-Bu;    -   Cys: Acm, Bzl, Bzl-OMe, BzI-Me, Trt;    -   Glu: Bzl, t-Bu;    -   Gln: Trt, Xan;    -   His: Boc, Dnp, Tos, Trt;    -   Lys: Boc, Z-CI, Fmoc, Z;    -   Ser: Bzl, TBDMS, TBDPS;    -   Thr: Bz;    -   Trp: Boc;    -   Tyr: Bzl, Z, Z-Br.

In one embodiment, —X₂— is connected indirectly to the Drug unit. Insuch an embodiment, the Spacer unit L² is present.

In one embodiment, the dipeptide is used in combination with aself-immolative group(s) (the Spacer unit). The self-immolative group(s)may be connected to —X₂—.

Where a self-immolative group is present, —X₂— is connected directly tothe self-immolative group. In one embodiment, —X₂— is connected to thegroup Y of the self-immolative group. Preferably the group —X₂—CO— isconnected to Y, where Y is NH.

—X₁— is connected directly to A¹. In one embodiment, —X¹— is connecteddirectly to A¹. Preferably the group NH—X₁— (the amino terminus of X₁)is connected to A¹. A¹ may comprise the functionality —CO— thereby toform an amide link with —X₁—.

In one embodiment, L¹ and L² together with —OC(═O)— comprise the group—X₁—X₂-PABC-. The PABC group is connected directly to the Drug unit. Inone example, the self-immolative group and the dipeptide together formthe group -Phe-Lys-PABC-, which is illustrated below:

where the asterisk indicates the point of attachment to the Drug unit,and the wavy line indicates the point of attachment to the remainingportion of L¹ or the point of attachment to A¹. Preferably, the wavyline indicates the point of attachment to A¹.

Alternatively, the self-immolative group and the dipeptide together formthe group -Val-Ala-PABC-, which is illustrated below:

-   -   where the asterisk and the wavy line are as defined above.

In another embodiment, L¹ and L² together with —OC(═O)— represent:

-   -   where the asterisk indicates the point of attachment to the Drug        unit, the wavy line indicates the point of attachment to A¹, Y        is a covalent bond or a functional group, and E is a group that        is susceptible to cleavage thereby to activate a self-immolative        group.

E is selected such that the group is susceptible to cleavage, e.g., bylight or by the action of an enzyme. E may be —NO₂ or glucuronic acid(e.g., β-glucuronic acid). The former may be susceptible to the actionof a nitroreductase, the latter to the action of a β-glucuronidase.

The group Y may be a covalent bond.

The group Y may be a functional group selected from:

-   -   —C(═O)—    -   —NH—    -   —O—    -   —C(═O)NH—,    -   —C(═O)O—,    -   —NHC(═O)—,    -   —OC(═O)—,    -   —OC(═O)O—,    -   —NHC(═O)O—,    -   —OC(═O)NH—,    -   —NHC(═O)NH—,    -   —NHC(═O)NH,    -   —C(═O)NHC(═O)—,    -   SO₂, and    -   —S—.

The group Y is preferably —NH—, —CH₂—, —O—, and —S—.

In some embodiments, L¹ and L² together with —OC(═O)— represent:

-   -   where the asterisk indicates the point of attachment to the Drug        unit, the wavy line indicates the point of attachment to A, Y is        a covalent bond or a functional group and E is glucuronic acid        (e.g., β-glucuronic acid). Y is preferably a functional group        selected from —NH—.

In some embodiments, L¹ and L² together represent:

-   -   where the asterisk indicates the point of attachment to the        remainder of L² or the Drug unit, the wavy line indicates the        point of attachment to A¹, Y is a covalent bond or a functional        group and E is glucuronic acid (e.g., β-glucuronic acid). Y is        preferably a functional group selected from —NH—, —CH₂—, —O—,        and —S—.

In some further embodiments, Y is a functional group as set forth above,the functional group is linked to an amino acid, and the amino acid islinked to the Stretcher unit A¹. In some embodiments, amino acid isβ-alanine. In such an embodiment, the amino acid is equivalentlyconsidered part of the Stretcher unit.

The Specificity unit L¹ and the Ligand unit are indirectly connected viathe Stretcher unit.

L¹ and A¹ may be connected by a bond selected from:

-   -   —C(═O)NH—,    -   —C(═O)O—,    -   —NHC(═O)—,    -   —OC(═O)—,    -   —OC(═O)O—,    -   —NHC(═O)O—,    -   —OC(═O)NH—, and    -   —NHC(═O)NH—.

A¹ may be of formula:-L^(A)-A²-whereinL^(A) is selected from:

where Ar represents a C₅₋₆ arylene group, e.g. phenylene.and A² is selected from:

-   -   n is 0 to 6, e.g. 5;

-   -   n is 0 to 6, e.g. 5;

-   -   n is 0 or 1 (e.g. 1), and m is 0 to 30 (preferably 0 to 10, 1 to        8, 4 to 8, most preferably 4 or 8);

-   -   n is 0 or 1 (e.g. 1), and m is 0 to 30 (preferably 0 to 10, 1 to        8, 4 to 8, most preferably 4 or 8);        where the asterisk indicates the point of attachment to L¹, the        wavy line indicates the point of attachment to L^(A).

In a particular embodiment, L^(A) is (L^(A1-1)) and A² is

where n is 1, and m is 0 to 8, e.g. 8

In one embodiment, the Stretcher unit A¹ is present, the Specificityunit L¹ is present and Spacer unit L² is absent. Thus, L¹ and the Drugunit are directly connected via a bond. Equivalently in this embodiment,L² is a bond.

L¹ and D may be connected by a bond selected from:

-   -   —C(═O)NH—,    -   —C(═O)O—,    -   —NHC(═O)—,    -   —OC(═O)—,    -   —OC(═O)O—,    -   —NHC(═O)O—,    -   —OC(═O)NH—, and    -   —NHC(═O)NH—.

In one embodiment, L¹ and D are preferably connected by a bond selectedfrom:

-   -   —C(═O)NH—, and    -   —NHC(═O)—.

In one embodiment, L¹ comprises a dipeptide and one end of the dipeptideis linked to D. As described above, the amino acids in the dipeptide maybe any combination of natural amino acids and non-natural amino acids.In some embodiments, the dipeptide comprises natural amino acids. Wherethe linker is a cathepsin labile linker, the dipeptide is the site ofaction for cathepsin-mediated cleavage. The dipeptide then is arecognition site for cathepsin.

In one embodiment, L¹-D is:

—NH—X₁-X₂—CO—NH—*

-   -   where —NH—X₁—X₂—CO is the dipeptide, —NH— is part of the Drug        unit, the asterisk indicates the point of attachment to the        remainder of the Drug unit, and the wavy line indicates the        point of attachment to the remaining portion of L¹ or the point        of attachment to A¹. Preferably, the wavy line indicates the        point of attachment to A¹.

In one embodiment, the dipeptide is valine-alanine and L¹-D is:

-   -   where the asterisk, —NH— and the wavy line are as defined above.

In one embodiment, the dipeptide is phenylalanine-lysine and L¹-D is:

-   -   where the asterisk, —NH— and the wavy line are as defined above.

In one embodiment, the dipeptide is valine-citrulline.

In other embodiments, Linker-Drug compounds are provided for conjugationto a Ligand unit. In one embodiment, the Linker-Drug compounds aredesigned for connection to a Cell Binding Agent.

In one embodiment, the Drug Linker unit has the formula:

-   -   where the asterisk indicates the point of attachment to the Drug        unit, G¹ is a Stretcher group (A¹) to form a connection to a        Ligand unit, L¹ is a Specificity unit, L² (a Spacer unit) is a        covalent bond or together with —OC(═O)— forms a self-immolative        group(s).

In another embodiment, the Drug Linker unit has the formula:G¹-L¹-L²-*where the asterisk indicates the point of attachment to the Drug unit,G¹ is a Stretcher unit (A¹) to form a connection to a Ligand unit, L¹ isa Specificity unit, L² (a Spacer unit) is a covalent bond or aself-immolative group(s).

In particular of these embodiments, the Drug Linker has the formula:G¹-L¹-

-   -   where the asterisk indicates the point of attachment to the Drug        unit, G¹ is a modified Stretcher unit (A¹) to form a connection        to a Ligand unit and L¹ is a Specificity unit.

L¹ and L² are as defined above. References to connection to A¹ can beconstrued here as referring to a connection to G¹.

The functional group G¹ forms a connecting group upon reaction with aLigand unit (e.g., a cell binding agent.

G¹ may be of formula:G^(A)-A²-where A² is as defined above and G^(A) is selected from:

where Ar represents a C₅₋₆ arylene group, e.g. phenylene.

In a particular embodiment, GA is (G^(A1-1)) and A² is

where n is 1, and m is 0 to 8, e.g. 8.

Ligand Unit

The Ligand Unit may be of any kind, and include a protein, polypeptide,peptide and a non-peptidic agent that specifically binds to a targetmolecule. In some embodiments, the Ligand unit may be a protein,polypeptide or peptide. In some embodiments, the Ligand unit may be acyclic polypeptide. These Ligand units can include antibodies or afragment of an antibody that contains at least one targetmolecule-binding site, lymphokines, hormones, growth factors, or anyother cell binding molecule or substance that can specifically bind to atarget.

The terms “specifically binds” and “specific binding” refer to thebinding of an antibody or other protein, polypeptide or peptide to apredetermined molecule (e.g., an antigen). Typically, the antibody orother molecule binds with an affinity of at least about 1×10⁷ M⁻¹, andbinds to the predetermined molecule with an affinity that is at leasttwo-fold greater than its affinity for binding to a non-specificmolecule (e.g., BSA, casein) other than the predetermined molecule or aclosely-related molecule.

Examples of Ligand units include those agents described for use in WO2007/085930, which is incorporated herein.

In some embodiments, the Ligand unit is a Cell Binding Agent that bindsto an extracellular target on a cell. Such a Cell Binding Agent can be aprotein, polypeptide, peptide or a non-peptidic agent. In someembodiments, the Cell Binding Agent may be a protein, polypeptide orpeptide. In some embodiments, the Cell Binding Agent may be a cyclicpolypeptide. The Cell Binding Agent also may be antibody or anantigen-binding fragment of an antibody. Thus, in one embodiment, thepresent invention provides an antibody-drug conjugate (ADC).

Cell Binding Agent

A cell binding agent may be of any kind, and include peptides andnon-peptides. These can include antibodies or a fragment of an antibodythat contains at least one binding site, lymphokines, hormones, hormonemimetics, vitamins, growth factors, nutrient-transport molecules, or anyother cell binding molecule or substance.

Peptides

In one embodiment, the cell binding agent is a linear or cyclic peptidecomprising 4-30, preferably 6-20, contiguous amino acid residues. Inthis embodiment, it is preferred that one cell binding agent is linkedto one monomer or dimer pyrrolobenzodiazepine compound.

In one embodiment the cell binding agent comprises a peptide that bindsintegrin α_(v)β₆. The peptide may be selective for α_(v)β₆ over XYS.

In one embodiment the cell binding agent comprises the A20FMDV-Cyspolypeptide. The A20FMDV-Cys has the sequence: NAVPNLRGDLQVLAQKVARTC.Alternatively, a variant of the A20FMDV-Cys sequence may be used whereinone, two, three, four, five, six, seven, eight, nine or ten amino acidresidues are substituted with another amino acid residue. Furthermore,the polypeptide may have the sequence

NAVXXXXXXXXXXXXXXXRTC.

Antibodies

The term “antibody” herein is used in the broadest sense andspecifically covers monoclonal antibodies, polyclonal antibodies,dimers, multimers, multispecific antibodies (e.g., bispecificantibodies), and antibody fragments, so long as they exhibit the desiredbiological activity (Miller et al (2003) Jour. of Immunology170:4854-4861). Antibodies may be murine, human, humanized, chimeric, orderived from other species. An antibody is a protein generated by theimmune system that is capable of recognizing and binding to a specificantigen. (Janeway, C., Travers, P., Walport, M., Shlomchik (2001) ImmunoBiology, 5th Ed., Garland Publishing, New York). A target antigengenerally has numerous binding sites, also called epitopes, recognizedby CDRs on multiple antibodies. Each antibody that specifically binds toa different epitope has a different structure. Thus, one antigen mayhave more than one corresponding antibody. An antibody includes afull-length immunoglobulin molecule or an immunologically active portionof a full-length immunoglobulin molecule, i.e., a molecule that containsan antigen binding site that immunospecifically binds an antigen of atarget of interest or part thereof, such targets including but notlimited to, cancer cell or cells that produce autoimmune antibodiesassociated with an autoimmune disease. The immunoglobulin can be of anytype (e.g. IgG, IgE, IgM, IgD, and IgA), class (e.g. IgG1, IgG2, IgG3,IgG4, IgA1 and IgA2) or subclass of immunoglobulin molecule. Theimmunoglobulins can be derived from any species, including human,murine, or rabbit origin.

“Antibody fragments” comprise a portion of a full length antibody,generally the antigen binding or variable region thereof. Examples ofantibody fragments include Fab, Fab′, F(ab′)₂, and scFv fragments;diabodies; linear antibodies; fragments produced by a Fab expressionlibrary, anti-idiotypic (anti-Id) antibodies, CDR (complementarydetermining region), and epitope-binding fragments of any of the abovewhich immunospecifically bind to cancer cell antigens, viral antigens ormicrobial antigens, single-chain antibody molecules; and multispecificantibodies formed from antibody fragments.

The term “monoclonal antibody” as used herein refers to an antibodyobtained from a population of substantially homogeneous antibodies, i.e.the individual antibodies comprising the population are identical exceptfor possible naturally occurring mutations that may be present in minoramounts. Monoclonal antibodies are highly specific, being directedagainst a single antigenic site. Furthermore, in contrast to polyclonalantibody preparations which include different antibodies directedagainst different determinants (epitopes), each monoclonal antibody isdirected against a single determinant on the antigen. In addition totheir specificity, the monoclonal antibodies are advantageous in thatthey may be synthesized uncontaminated by other antibodies. The modifier“monoclonal” indicates the character of the antibody as being obtainedfrom a substantially homogeneous population of antibodies, and is not tobe construed as requiring production of the antibody by any particularmethod. For example, the monoclonal antibodies to be used in accordancewith the present invention may be made by the hybridoma method firstdescribed by Kohler et al (1975) Nature 256:495, or may be made byrecombinant DNA methods (see, U.S. Pat. No. 4,816,567). The monoclonalantibodies may also be isolated from phage antibody libraries using thetechniques described in Clackson et al (1991) Nature, 352:624-628; Markset al (1991) J. Mol. Biol., 222:581-597 or from transgenic mice carryinga fully human immunoglobulin system (Lonberg (2008) Curr. Opinion20(4):450-459).

The monoclonal antibodies herein specifically include “chimeric”antibodies in which a portion of the heavy and/or light chain isidentical with or homologous to corresponding sequences in antibodiesderived from a particular species or belonging to a particular antibodyclass or subclass, while the remainder of the chain(s) is identical withor homologous to corresponding sequences in antibodies derived fromanother species or belonging to another antibody class or subclass, aswell as fragments of such antibodies, so long as they exhibit thedesired biological activity (U.S. Pat. No. 4,816,567; and Morrison et al(1984) Proc. Natl. Acad. Sci. USA, 81:6851-6855). Chimeric antibodiesinclude “primatized” antibodies comprising variable domainantigen-binding sequences derived from a non-human primate (e.g. OldWorld Monkey or Ape) and human constant region sequences.

An “intact antibody” herein is one comprising a VL and VH domains, aswell as a light chain constant domain (CL) and heavy chain constantdomains, CH1, CH2 and CH3. The constant domains may be native sequenceconstant domains (e.g. human native sequence constant domains) or aminoacid sequence variant thereof. The intact antibody may have one or more“effector functions” which refer to those biological activitiesattributable to the Fc region (a native sequence Fc region or amino acidsequence variant Fc region) of an antibody. Examples of antibodyeffector functions include C1q binding; complement dependentcytotoxicity; Fc receptor binding; antibody-dependent cell-mediatedcytotoxicity (ADCC); phagocytosis; and down regulation of cell surfacereceptors such as B cell receptor and BCR.

Depending on the amino acid sequence of the constant domain of theirheavy chains, intact antibodies can be assigned to different “classes.”There are five major classes of intact antibodies: IgA, IgD, IgE, IgG,and IgM, and several of these may be further divided into “subclasses”(isotypes), e.g., IgG1, IgG2, IgG3, IgG4, IgA, and IgA2. The heavy-chainconstant domains that correspond to the different classes of antibodiesare called α, δ, ε, γ, and μ, respectively. The subunit structures andthree-dimensional configurations of different classes of immunoglobulinsare well known.

Humanisation

Techniques to reduce the in vivo immunogenicity of a non-human antibodyor antibody fragment include those termed “humanisation”.

A “humanized antibody” refers to a polypeptide comprising at least aportion of a modified variable region of a human antibody wherein aportion of the variable region, preferably a portion substantially lessthan the intact human variable domain, has been substituted by thecorresponding sequence from a non-human species and wherein the modifiedvariable region is linked to at least another part of another protein,preferably the constant region of a human antibody. The expression“humanized antibodies” includes human antibodies in which one or morecomplementarity determining region (“CDR”) amino acid residues and/orone or more framework region (“FW” or “FR”) amino acid residues aresubstituted by amino acid residues from analogous sites in rodent orother non-human antibodies. The expression “humanized antibody” alsoincludes an immunoglobulin amino acid sequence variant or fragmentthereof that comprises an FR having substantially the amino acidsequence of a human immunoglobulin and a CDR having substantially theamino acid sequence of a non-human immunoglobulin.

“Humanized” forms of non-human (e.g., murine) antibodies are chimericantibodies that contain minimal sequence derived from non-humanimmunoglobulin. Or, looked at another way, a humanized antibody is ahuman antibody that also contains selected sequences from non-human(e.g. murine) antibodies in place of the human sequences. A humanizedantibody can include conservative amino acid substitutions ornon-natural residues from the same or different species that do notsignificantly alter its binding and/or biologic activity. Suchantibodies are chimeric antibodies that contain minimal sequence derivedfrom non-human immunoglobulins.

There are a range of humanisation techniques, including ‘CDR grafting’,‘guided selection’, ‘deimmunization’, ‘resurfacing’ (also known as‘veneering), ‘composite antibodies’, ‘Human String Content Optimisation’and framework shuffling.

CDR Grafting

In this technique, the humanized antibodies are human immunoglobulins(recipient antibody) in which residues from a complementary-determiningregion (CDR) of the recipient antibody are replaced by residues from aCDR of a non-human species (donor antibody) such as mouse, rat, camel,bovine, goat, or rabbit having the desired properties (in effect, thenon-human CDRs are ‘grafted’ onto the human framework). In someinstances, framework region (FR) residues of the human immunoglobulinare replaced by corresponding non-human residues (this may happen when,for example, a particular FR residue has significant effect on antigenbinding).

Furthermore, humanized antibodies can comprise residues that are foundneither in the recipient antibody nor in the imported CDR or frameworksequences. These modifications are made to further refine and maximizeantibody performance. Thus, in general, a humanized antibody willcomprise all of at least one, and in one aspect two, variable domains,in which all or all of the hypervariable loops correspond to those of anon-human immunoglobulin and all or substantially all of the FR regionsare those of a human immunoglobulin sequence. The humanized antibodyoptionally also will comprise at least a portion of an immunoglobulinconstant region (Fc), or that of a human immunoglobulin.

Guided Selection

The method consists of combining the V_(H) or V_(L) domain of a givennon-human antibody specific for a particular epitope with a human V_(H)or V_(L) library and specific human V domains are selected against theantigen of interest. This selected human VH is then combined with a VLlibrary to generate a completely human VH×VL combination. The method isdescribed in Nature Biotechnology (N.Y.) 12, (1994) 899-903.

Composite Antibodies

In this method, two or more segments of amino acid sequence from a humanantibody are combined within the final antibody molecule. They areconstructed by combining multiple human VH and VL sequence segments incombinations which limit or avoid human T cell epitopes in the finalcomposite antibody V regions. Where required, T cell epitopes arelimited or avoided by, exchanging V region segments contributing to orencoding a T cell epitope with alternative segments which avoid T cellepitopes. This method is described in US 2008/0206239 A1.

Deimmunization

This method involves the removal of human (or other second species)T-cell epitopes from the V regions of the therapeutic antibody (or othermolecule). The therapeutic antibodies V-region sequence is analysed forthe presence of MHC class II-binding motifs by, for example, comparisonwith databases of MHC-binding motifs (such as the “motifs” databasehosted at www.wehi.edu.au). Alternatively, MHC class II-binding motifsmay be identified using computational threading methods such as thosedevised by Altuvia et al. (J. Mol. Biol. 249 244-250 (1995)); in thesemethods, consecutive overlapping peptides from the V-region sequencesare testing for their binding energies to MHC class II proteins. Thisdata can then be combined with information on other sequence featureswhich relate to successfully presented peptides, such as amphipathicity,Rothbard motifs, and cleavage sites for cathepsin B and other processingenzymes.

Once potential second species (e.g. human) T-cell epitopes have beenidentified, they are eliminated by the alteration of one or more aminoacids. The modified amino acids are usually within the T-cell epitopeitself, but may also be adjacent to the epitope in terms of the primaryor secondary structure of the protein (and therefore, may not beadjacent in the primary structure). Most typically, the alteration is byway of substitution but, in some circumstances amino acid addition ordeletion will be more appropriate.

All alterations can be accomplished by recombinant DNA technology, sothat the final molecule may be prepared by expression from a recombinanthost using well established methods such as Site Directed Mutagenesis.However, the use of protein chemistry or any other means of molecularalteration is also possible.

Resurfacing

This method involves:

-   -   (a) determining the conformational structure of the variable        region of the non-human (e.g. rodent) antibody (or fragment        thereof) by constructing a three-dimensional model of the        non-human antibody variable region;    -   (b) generating sequence alignments using relative accessibility        distributions from x-ray crystallographic structures of a        sufficient number of non-human and human antibody variable        region heavy and light chains to give a set of heavy and light        chain framework positions wherein the alignment positions are        identical in 98% of the sufficient number of non-human antibody        heavy and light chains;    -   (c) defining for the non-human antibody to be humanized, a set        of heavy and light chain surface exposed amino acid residues        using the set of framework positions generated in step (b);    -   (d) identifying from human antibody amino acid sequences a set        of heavy and light chain surface exposed amino acid residues        that is most closely identical to the set of surface exposed        amino acid residues defined in step (c), wherein the heavy and        light chain from the human antibody are or are not naturally        paired;    -   (e) substituting, in the amino acid sequence of the non-human        antibody to be humanized, the set of heavy and light chain        surface exposed amino acid residues defined in step (c) with the        set of heavy and light chain surface exposed amino acid residues        identified in step (d);    -   (f) constructing a three-dimensional model of the variable        region of the non-human antibody resulting from the substituting        specified in step (e);    -   (g) identifying, by comparing the three-dimensional models        constructed in steps (a) and (f), any amino acid residues from        the sets identified in steps (c) or (d), that are within 5        Angstroms of any atom of any residue of the complementarity        determining regions of the non-human antibody to be humanized;        and    -   (h) changing any residues identified in step (g) from the human        to the original non-human amino acid residue to thereby define a        non-human antibody humanizing set of surface exposed amino acid        residues; with the proviso that step (a) need not be conducted        first, but must be conducted prior to step (g).

Superhumanization

The method compares the non-human sequence with the functional humangermline gene repertoire. Those human genes encoding canonicalstructures identical or closely related to the non-human sequences areselected. Those selected human genes with highest homology within theCDRs are chosen as FR donors. Finally, the non-human CDRs are graftedonto these human FRs. This method is described in patent WO 2005/079479A2.

Human String Content Optimization

This method compares the non-human (e.g. mouse) sequence with therepertoire of human germline genes and the differences are scored asHuman String Content (HSC) that quantifies a sequence at the level ofpotential MHC/T-cell epitopes. The target sequence is then humanized bymaximizing its HSC rather than using a global identity measure togenerate multiple diverse humanized variants (described in MolecularImmunology, 44, (2007) 1986-1998).

Framework Shuffling

The CDRs of the non-human antibody are fused in-frame to cDNA poolsencompassing all known heavy and light chain human germline geneframeworks. Humanised antibodies are then selected by e.g. panning ofthe phage displayed antibody library. This is described in Methods 36,43-60 (2005).

Examples of cell binding agents include those agents described for usein WO 2007/085930, which is incorporated herein.

Tumour-associate antigens and cognate antibodies for use in embodimentsof the present invention are listed below.

Tumor-Associated Antigens and Cognate Antibodies

(1) BMPR1B (Bone Morphogenetic Protein Receptor-Type IB)

Nucleotide

Genbank accession no. NM_001203

Genbank version no. NM_001203.2 GI:169790809

Genbank record update date: Sep. 23, 2012 02:06 PM

Polypeptide

Genbank accession no. NP_001194

Genbank version no. NP_001194.1 GI:4502431

Genbank record update date: Sep. 23, 2012 02:06 PM

Cross-References

ten Dijke, P., et al Science 264 (5155): 101-104 (1994), Oncogene 14(11):1377-1382 (1997)); WO2004/063362 (claim 2); WO2003/042661 (claim12); US2003/134790-A1 (Page 38-39); WO2002/102235 (claim 13; Page 296);WO2003/055443 (Page 91-92); WO2002/99122 (Example 2; Page 528-530);WO2003/029421 (claim 6); WO2003/024392 (claim 2; FIG. 112); WO2002/98358(claim 1; Page 183); WO2002/54940 (Page 100-101); WO2002/59377(Page349-350); WO2002/30268 (claim 27; Page 376); WO2001/48204 (Example; FIG.4); NP_001194 bone morphogenetic protein receptor, type IB/pid=NP_001194.1.; MIM:603248; AY065994

(2) E16 (LAT1, SLC7A5)

Nucleotide

Genbank accession no. NM_003486

Genbank version no. NM_003486.5 GI:71979931

Genbank record update date: Jun. 27, 2012 12:06 PM

Polypeptide

Genbank accession no. NP_003477

Genbank version no. NP_003477.4 GI:71979932

Genbank record update date: Jun. 27, 2012 12:06 PM

Cross References

Biochem. Biophys. Res. Commun. 255 (2), 283-288 (1999), Nature 395(6699):288-291 (1998), Gaugitsch, H. W., et al (1992) J. Biol. Chem. 267(16):11267-11273); WO2004/048938 (Example 2); WO2004/032842 (ExampleIV); WO2003/042661 (claim 12); WO2003/016475 (claim 1); WO2002/78524(Example 2); WO2002/99074 (claim 19; Page 127-129); WO2002/86443 (claim27; Pages 222, 393); WO2003/003906 (claim 10; Page 293); WO2002/64798(claim 33; Page 93-95); WO2000/14228 (claim 5; Page 133-136);US2003/224454 (FIG. 3); WO2003/025138 (claim 12; Page 150); NP_003477solute carrier family 7 (cationic amino acid transporter, y+system),member 5/pid=NP_003477.3—Homo sapiens; MIM:600182; NM_015923.

(3) STEAP1 (Six Transmembrane Epithelial Antigen of Prostate)

Nucleotide

Genbank accession no. NM_012449

Genbank version no. NM_012449.2 GI:22027487

Genbank record update date: Sep. 9, 2012 02:57 PM

Polypeptide

Genbank accession no. NP_036581

Genbank version no. NP_036581.1 GI:9558759

Genbank record update date: Sep. 9, 2012 02:57 PM

Cross References

Cancer Res. 61 (15), 5857-5860 (2001), Hubert, R. S., et al (1999) Proc.Natl. Acad. Sci. U.S.A. 96 (25): 14523-14528); WO2004/065577 (claim 6);WO2004/027049 (FIG. 1L); EP1394274 (Example 11); WO2004/016225 (claim2); WO2003/042661 (claim 12); US2003/157089 (Example 5); US2003/185830(Example 5); US2003/064397 (FIG. 2); WO2002/89747 (Example 5; Page618-619); WO2003/022995 (Example 9; FIG. 13A, Example 53; Page 173,Example 2; FIG. 2A); six transmembrane epithelial antigen of theprostate; MIM:604415.

(4) 0772P (CA125, MUC16)

Nucleotide

Genbank accession no. AF361486

Genbank version no. AF361486.3 GI:34501466

Genbank record update date: Mar. 11, 2010 07:56 AM

Polypeptide

Genbank accession no. AAK74120

Genbank version no. AAK74120.3 GI:34501467

Genbank record update date: Mar. 11, 2010 07:56 AM

Cross References

J. Biol. Chem. 276 (29):27371-27375 (2001)); WO2004/045553 (claim 14);WO2002/92836 (claim 6; FIG. 12); WO2002/83866 (claim 15; Page 116-121);US2003/124140 (Example 16); GI:34501467;

(5) MPF (MPF, MSLN, SMR, Megakaryocyte Potentiating Factor, Mesothelin)

Nucleotide

Genbank accession no. NM_005823

Genbank version no. NM_005823.5 GI:293651528

Genbank record update date: Sep. 2, 2012 01:47 PM

Polypeptide

Genbank accession no. NP_005814

Genbank version no. NP_005814.2 GI:53988378

Genbank record update date: Sep. 2, 2012 01:47 PM

Cross References

Yamaguchi, N., et al Biol. Chem. 269 (2), 805-808 (1994), Proc. Natl.Acad. Sci. U.S.A. 96 (20):11531-11536 (1999), Proc. Natl. Acad. Sci.U.S.A. 93 (1):136-140 (1996), J. Biol. Chem. 270 (37):21984-21990(1995)); WO2003/101283 (claim 14); (WO2002/102235 (claim 13; Page287-288); WO2002/101075 (claim 4; Page 308-309); WO2002/71928 (Page320-321); WO94/10312 (Page 52-57); IM:601051.

(6) Napi3b (NAPI-3B, NPTIIb, SLC34A2, Solute Carrier Family 34 (SodiumPhosphate), Member 2, Type II Sodium-Dependent Phosphate Transporter 3b)

Nucleotide

Genbank accession no. NM_006424

Genbank version no. NM_006424.2 GI:110611905

Genbank record update date: Jul. 22, 2012 03:39 PM

Polypeptide

Genbank accession no. NP_006415

Genbank version no. NP_006415.2 GI:110611906

Genbank record update date: Jul. 22, 2012 03:39 PM

Cross References

J. Biol. Chem. 277 (22):19665-19672 (2002), Genomics 62 (2):281-284(1999), Feild, J. A., et al (1999) Biochem. Biophys. Res. Commun. 258(3):578-582); WO2004/022778 (claim 2); EP1394274 (Example 11);WO2002/102235 (claim 13; Page 326); EP0875569 (claim 1; Page 17-19);WO2001/57188 (claim 20; Page 329); WO2004/032842 (Example IV);WO2001/75177 (claim 24; Page 139-140); MIM:604217.

(7) Sema 5b (FLJ10372, KIAA1445, Mm.42015, SEMA5B, SEMAG, Semaphorin 5bHlog, 25 sema domain, seven thrombospondin repeats (type 1 and type1-like), transmembrane domain (TM) and short cytoplasmic domain,(semaphorin) 58)

Nucleotide

Genbank accession no. AB040878

Genbank version no. AB040878.1 GI:7959148

Genbank record update date: Aug. 2, 2006 05:40 PM

Polypeptide

Genbank accession no. BAA95969

Genbank version no. BAA95969.1 GI:7959149

Genbank record update date: Aug. 2, 2006 05:40 PM

Cross References

Nagase T., et al (2000) DNA Res. 7 (2):143-150); WO2004/000997 (claim1); WO2003/003984 (claim 1); WO2002/06339 (claim 1; Page 50);WO2001/88133 (claim 1; Page 41-43, 48-58); WO2003/054152 (claim 20);WO2003/101400 (claim 11); Accession: Q9P283; Genew; HGNC:10737

(8) PSCA hlg (2700050C12Rik, C530008O16Rik, RIKEN cDNA 2700050C12, RIKENcDNA 2700050C12 gene)

Nucleotide

Genbank accession no. AY358628

Genbank version no. AY358628.1 GI:37182377

Genbank record update date: Dec. 1, 2009 04:15 AM

Polypeptide

Genbank accession no. AAQ88991

Genbank version no. AAQ88991.1 GI:37182378

Genbank record update date: Dec. 1, 2009 04:15 AM

Cross References

Ross et al (2002) Cancer Res. 62:2546-2553; US2003/129192 (claim 2);US2004/044180 (claim 12); US2004/044179 (claim 11); US2003/096961 (claim11); US2003/232056 (Example 5); WO2003/105758 16 (claim 12);US2003/206918 (Example 5); EP1347046 (claim 1); WO2003/025148 (claim20); GI:37182378.

(9) ETBR (Endothelin type B receptor)

Nucleotide

Genbank accession no. AY275463

Genbank version no. AY275463.1 GI:30526094

Genbank record update date: Mar. 11, 2010 02:26 AM

Polypeptide

Genbank accession no. AAP32295

Genbank version no. AAP32295.1 GI:30526095

Genbank record update date: Mar. 11, 2010 02:26 AM

Cross References

Nakamuta M., et al Biochem. Biophys. Res. Commun. 177, 34-39, 1991;Ogawa Y., et al Biochem. Biophys. Res. Commun. 178, 248-255, 1991; AraiH., et al Jpn. Circ. J. 56, 1303-1307, 1992; Arai H., et al J. Biol.Chem. 268, 3463-3470, 1993; Sakamoto A., Yanagisawa M., et al Biochem.Biophys. Res. Commun. 178, 656-663, 1991; Elshourbagy N. A., et al J.Biol. Chem. 268, 3873-3879, 1993; Haendler B., et al J. Cardiovasc.Pharmacol. 20, s1-S4, 1992; Tsutsumi M., et al Gene 228, 43-49, 1999;Strausberg R. L., et al Proc. Natl. Acad. Sci. U.S.A. 99, 16899-16903,2002; Bourgeois C., et al J. Clin. Endocrinol. Metab. 82, 3116-3123,1997; Okamoto Y., et al Biol. Chem. 272, 21589-21596, 1997; Verheij J.B., et al Am. J. Med. Genet. 108, 223-225, 2002; Hofstra R. M. W., et alEur. J. Hum. Genet. 5, 180-185, 1997; Puffenberger E. G., et al Cell 79,1257-1266, 1994; Attie T., et al, Hum. Mol. Genet. 4, 2407-2409, 1995;Auricchio A., et al Hum. Mol. Genet. 5:351-354, 1996; Amiel J., et alHum. Mol. Genet. 5, 355-357, 1996; Hofstra R. M. W., et al Nat. Genet.12, 445-447, 1996; Svensson P. J., et al Hum. Genet. 103, 145-148, 1998;Fuchs S., et al Mol. Med. 7, 115-124, 2001; Pingault V., et al (2002)Hum. Genet. 111, 198-206; WO2004/045516 (claim 1); WO2004/048938(Example 2); WO2004/040000 (claim 151); WO2003/087768 (claim 1);WO2003/016475 (claim 1); WO2003/016475 (claim 1); WO2002/61087 (FIG. 1);WO2003/016494 (FIG. 6); WO2003/025138 (claim 12; Page 144); WO2001/98351(claim 1; Page 124-125); EP0522868 (claim 8; FIG. 2); WO2001/77172(claim 1; Page 297-299); US2003/109676; U.S. Pat. No. 6,518,404 (FIG.3); U.S. Pat. No. 5,773,223 (Claim 1a; Col 31-34); WO2004/001004.

(10) MSG783 (RNF124, hypothetical protein FLJ20315)

Nucleotide

Genbank accession no. NM_017763

Genbank version no. NM_017763.4 GI:167830482

Genbank record update date: Jul. 22, 2012 12:34 AM

Polypeptide

Genbank accession no. NP_060233

Genbank version no. NP_060233.3 GI:56711322

Genbank record update date: Jul. 22, 2012 12:34 AM

Cross References

WO2003/104275 (claim 1); WO2004/046342 (Example 2); WO2003/042661 (claim12); WO2003/083074 (claim 14; Page 61); WO2003/018621 (claim 1);WO2003/024392 (claim 2; FIG. 93); WO2001/66689 (Example 6);LocusID:54894.

(11) STEAP2 (HGNC 8639, IPCA-1, PCANAP1, STAMP1, STEAP2, STMP, prostatecancer

associated gene 1, prostate cancer associated protein 1, sixtransmembrane epithelial antigen of prostate 2, six transmembraneprostate protein)

Nucleotide

Genbank accession no. AF455138

Genbank version no. AF455138.1 GI:22655487

Genbank record update date: Mar. 11, 2010 01:54 AM

Polypeptide

Genbank accession no. AAN04080

Genbank version no. AAN04080.1 GI:22655488

Genbank record update date: Mar. 11, 2010 01:54 AM

Cross References

Lab. Invest. 82 (11):1573-1582 (2002)); WO2003/087306; US2003/064397(claim 1; FIG. 1); WO2002/72596 (claim 13; Page 54-55); WO2001/72962(claim 1; FIG. 4B); WO2003/104270 (claim 11); WO2003/104270 (claim 16);US2004/005598 (claim 22); WO2003/042661 (claim 12); US2003/060612 (claim12; FIG. 10); WO2002/26822 (claim 23; FIG. 2); WO2002/16429 (claim 12;FIG. 10); GI:22655488.

(12) TrpM4 (BR22450, FLJ20041, TRPM4, TRPM4B, transient receptorpotential cation channel, subfamily M, member 4)

Nucleotide

Genbank accession no. NM_017636

Genbank version no. NM_017636.3 GI:304766649

Genbank record update date: Jun. 29, 2012 11:27 AM

Polypeptide

Genbank accession no. NP_060106

Genbank version no. NP_060106.2 GI:21314671

Genbank record update date: Jun. 29, 2012 11:27 AM

Cross References

Xu, X. Z., et al Proc. Natl. Acad. Sci. U.S.A. 98 (19):10692-10697(2001), Cell 109 (3):397-407 (2002), J. Biol. Chem. 278 (33):30813-30820(2003)); US2003/143557 (claim 4); WO2000/40614 (claim 14; Page 100-103);WO2002/10382 (claim 1; FIG. 9A); WO2003/042661 (claim 12); WO2002/30268(claim 27; Page 391); US2003/219806 (claim 4); WO2001/62794 (claim 1014; FIG. 1A-D); MIM:606936.

(13) CRIPTO (CR, CR1, CRGF, CRIPTO, TDGF1, teratocarcinoma-derivedgrowth factor)

Nucleotide

Genbank accession no. NM_003212

Genbank version no. NM_003212.3 GI:292494881

Genbank record update date: Sep. 23, 2012 02:27 PM

Polypeptide

Genbank accession no. NP_003203

Genbank version no. NP_003203.1 GI:4507425

Genbank record update date: Sep. 23, 2012 02:27 PM

Cross References

Ciccodicola, A., et al EMBO J. 8 (7):1987-1991 (1989), Am. J. Hum.Genet. 49 (3):555-565 (1991)); US2003/224411 (claim 1); WO2003/083041(Example 1); WO2003/034984 (claim 12); WO2002/88170 (claim 2; Page52-53); WO2003/024392 (claim 2; FIG. 58); WO2002/16413 (claim 1; Page94-95, 105); WO2002/22808 (claim 2; FIG. 1); U.S. Pat. No. 5,854,399(Example 2; Col 17-18); U.S. Pat. No. 5,792,616 (FIG. 2); MIM:187395.

(14) CD21 (CR2 (Complement receptor 2) or C3DR (C3d/Epstein Barr virusreceptor) or Hs. 73792)

Nucleotide

Genbank accession no M26004

Genbank version no. M26004.1 GI:181939

Genbank record update date: Jun. 23, 2010 08:47 AM

Polypeptide

Genbank accession no. AAA35786

Genbank version no. AAA35786.1 GI:181940

Genbank record update date: Jun. 23, 2010 08:47 AM

Cross References

Fujisaku et al (1989) J. Biol. Chem. 264 (4):2118-2125); Weis J. J., etal J. Exp. Med. 167, 1047-1066, 1988; Moore M., et al Proc. Natl. Acad.Sci. U.S.A. 84, 9194-9198, 1987; Barel M., et al Mol. Immunol. 35,1025-1031, 1998; Weis J. J., et al Proc. Natl. Acad. Sci. U.S.A. 83,5639-5643, 1986; Sinha S. K., et al (1993) J. Immunol. 150, 5311-5320;WO2004/045520 (Example 4); US2004/005538 (Example 1); WO2003/062401(claim 9); WO2004/045520 (Example 4); WO91/02536 (FIGS. 9.1-9.9);WO2004/020595 (claim 1); Accession: P20023; Q13866; Q14212; EMBL;M26004; AAA35786.1.

(15) CD79b (CD79B, CD79β, IGb (immunoglobulin-associated beta), 829)

Nucleotide

Genbank accession no NM_000626

Genbank version no. NM_000626.2 GI:90193589

Genbank record update date: Jun. 26, 2012 01:53 PM

Polypeptide

Genbank accession no. NP_000617

Genbank version no. NP_000617.1 GI:11038674

Genbank record update date: Jun. 26, 2012 01:53 PM

Cross References

Proc. Natl. Acad. Sci. U.S.A. (2003) 100 (7):4126-4131, Blood (2002) 100(9):3068-3076, Muller et al (1992) Eur. J. Immunol. 22 (6):1621-1625);WO2004/016225 (claim 2, FIG. 140); WO2003/087768, US2004/101874 (claim1, page 102); WO2003/062401 (claim 9); WO2002/78524 (Example 2);US2002/150573 (claim 5, page 15); U.S. Pat. No. 5,644,033; WO2003/048202(claim 1, pages 306 and 309); WO 99/58658, U.S. Pat. No. 6,534,482(claim 13, FIG. 17A/B); WO2000/55351 (claim 11, pages 1145-1146);MIM:147245

(16) FcRH2 (IFGP4, IRTA4, SPAP1A (SH2 domain containing phosphataseanchor protein 1a), SPAP1B, SPAP1C)

Nucleotide

Genbank accession no NM_030764

Genbank version no. NM_030764.3 GI:227430280

Genbank record update date: Jun. 30, 2012 12:30 AM

Polypeptide

Genbank accession no. NP_110391

Genbank version no. NP_110391.2 GI:19923629

Genbank record update date: Jun. 30, 2012 12:30 AM

Cross References

AY358130); Genome Res. 13 (10):2265-2270 (2003), Immunogenetics 54(2):87-95 (2002), Blood 99 (8):2662-2669 (2002), Proc. Natl. Acad. Sci.U.S.A. 98 (17):9772-9777 (2001), Xu, M. J., et al (2001) Biochem.Biophys. Res. Commun. 280 (3):768-775; WO2004/016225 (claim 2);WO2003/077836; WO2001/38490 (claim 5; FIG. 18D-1-18D-2); WO2003/097803(claim 12); WO2003/089624 (claim 25); MIM:606509.

(17) HER2 (ErbB2)

Nucleotide

Genbank accession no M11730

Genbank version no. M11730.1 GI:183986

Genbank record update date: Jun. 23, 2010 08:47 AM

Polypeptide

Genbank accession no. AAA75493

Genbank version no. AAA75493.1 GI:306840

Genbank record update date: Jun. 23, 2010 08:47 AM

Cross References

Coussens L., et al Science (1985) 230(4730):1132-1139); Yamamoto T., etal Nature 319, 230-234, 1986; Semba K., et al Proc. Natl. Acad. Sci.U.S.A. 82, 6497-6501, 1985; Swiercz J. M., et al J. Cell Biol. 165,869-15 880, 2004; Kuhns J. J., et al J. Biol. Chem. 274, 36422-36427,1999; Cho H.-S., et al Nature 421, 756-760, 2003; Ehsani A., et al(1993) Genomics 15, 426-429; WO2004/048938 (Example 2); WO2004/027049(FIG. 11); WO2004/009622; WO2003/081210; WO2003/089904 (claim 9);WO2003/016475 (claim 1); US2003/118592; WO2003/008537 (claim 1);WO2003/055439 (claim 29; FIG. 1A-B); WO2003/025228 (claim 37; FIG. 5C);WO2002/22636 (Example 13; Page 95-107); WO2002/12341 (claim 68; FIG. 7);WO2002/13847 (Page 71-74); WO2002/14503 (Page 114-117); WO2001/53463(claim 2; Page 41-46); WO2001/41787 (Page 15); WO2000/44899 (claim 52;FIG. 7); WO2000/20579 (claim 3; FIG. 2); U.S. Pat. No. 5,869,445 (claim3; Col 31-38); WO9630514 (claim 2; Page 56-61); EP1439393 (claim 7);WO2004/043361 (claim 7); WO2004/022709; WO2001/00244 (Example 3; FIG.4); Accession: P04626; EMBL; M11767; AAA35808.1. EMBL; M11761;AAA35808.1

Antibodies

Abbott: US20110177095

-   -   For example, an antibody comprising CDRs having overall at least        80% sequence identity to CDRs having amino acid sequences of SEQ        ID NO:3 (CDR-H1), SEQ ID NO:4 (CDR-H2), SEQ ID NO:5 (CDR-H3),        SEQ ID NO:104 and/or SEQ ID NO:6 (CDR-L1), SEQ ID NO:7 (CDR-L2),        and SEQ ID NO:8 (CDR-L3), wherein the anti-HER2 antibody or        anti-HER2 binding fragment has reduced immunogenicity as        compared to an antibody having a VH of SEQ ID NO:1 and a VL of        SEQ ID NO:2.

Biogen: US20100119511

-   -   For example, ATCC accession numbers: PTA-10355, PTA-10356,        PTA-10357, PTA-10358    -   For example, a purified antibody molecule that binds to HER2        comprising a all six CDR's from an antibody selected from the        group consisting of BIIB71F10 (SEQ ID NOs:11, 13), BIIB69A09        (SEQ ID NOs:15, 17); BIIB67F10 (SEQ ID NOs:19, 21); BIIB67F11        (SEQ ID NOs:23, 25), BIIB66A12 (SEQ ID NOs:27, 29), BIIB66C01        (SEQ ID NOs:31, 33), BIIB65C10 (SEQ ID NOs:35, 37), BIIB65H09        (SEQ ID NOs:39, 41) and 811665603 (SEQ ID NOs:43, 45), or CDRs        which are identical or which have no more than two alterations        from said CDRs.

Herceptin (Genentech)—U.S. Pat. No. 6,054,297; ATCC accession no.CRL-10463 (Genentech)

Pertuzumab (Genentech)

-   -   US20110117097        -   for example, see SEQ IDs No. 15&16, SEQ IDs No. 17&18, SEQ            IDs No. 23&24 & ATCC accession numbers HB-12215, HB-12216,            CRL 10463, HB-12697.    -   US20090285837    -   US20090202546        -   for example, ATCC accession numbers: HB-12215, HB-12216, CRL            10463, HB-12698.    -   US20060088523        -   for example, ATCC accession numbers: HB-12215, HB-12216        -   for example, an antibody comprising the variable light and            variable heavy amino acid sequences in SEQ ID Nos. 3 and 4,            respectively.        -   for example, an antibody comprising a light chain amino acid            sequence selected from SEQ ID No. 15 and 23, and a heavy            chain amino acid sequence selected from SEQ ID No. 16 and 24    -   US20060018899        -   for example, ATCC accession numbers: (7C2) HB-12215, (7F3)            HB-12216, (4D5) CRL-10463, (2C4) HB-12697.        -   for example, an antibody comprising the amino acid sequence            in SEQ ID No. 23, or a deamidated and/or oxidized variant            thereof.    -   US2011/0159014        -   for example, an antibody having a light chain variable            domain comprising the hypervariable regions of SEQ ID NO:            1″.        -   For example, an antibody having a heavy chain variable            domain comprising the hypervariable regions of SEQ ID NO: 2.    -   US20090187007

Glycotope: TrasGEX antibody http://www.glycotope.com/pipeline

-   -   For example, see International Joint Cancer Institute and        Changhai Hospital Cancer Cent: HMTI-Fc Ab-Gao J., et al BMB Rep.        2009 Oct. 31; 42(10):636-41.

Symphogen: US20110217305

Union Stem Cell & Gene Engineering, China-Liu H Q., et al Xi Bao Yu FenZi Mian Yi Xue Za Zhi. 2010 May; 26(5):456-8.

(18) NCA (CEACAM6)

Nucleotide

Genbank accession no M18728

Genbank version no. M18728.1 GI:189084

Genbank record update date: Jun. 23, 2010 08:48 AM

Polypeptide

Genbank accession no. AAA59907

Genbank version no. AAA59907.1 GI:189085

Genbank record update date: Jun. 23, 2010 08:48 AM

Cross References

Barnett T., et al Genomics 3, 59-66, 1988; Tawaragi Y., et al Biochem.Biophys. Res. Commun. 150, 89-96, 1988; Strausberg R. L., et al Proc.Natl. Acad. Sci. U.S.A. 99:16899-16903, 2002; WO2004/063709; EP1439393(claim 7); WO2004/044178 (Example 4); WO2004/031238; WO2003/042661(claim 12); WO2002/78524 (Example 2); WO2002/86443 (claim 27; Page 427);WO2002/60317 (claim 2); Accession: P40199; Q14920; EMBL; M29541;AAA59915.1. EMBL; M18728.

(19) MDP (DPEP1)

Nucleotide

Genbank accession no BC017023

Genbank version no. BC017023.1 GI:16877538

Genbank record update date: Mar. 6, 2012 01:00 PM

Polypeptide

Genbank accession no. AAH17023

Genbank version no. AAH17023.1 GI:16877539

Genbank record update date: Mar. 6, 2012 01:00 PM

Cross References

Proc. Natl. Acad. Sci. U.S.A. 99 (26):16899-16903 (2002)); WO2003/016475(claim 1); WO2002/64798 (claim 33; Page 85-87); JP05003790 (FIG. 6-8);WO99/46284 (FIG. 9); MIM:179780.

(20) IL20R-alpha (11_20Ra, ZCYTOR7)

Nucleotide

Genbank accession no AF184971

Genbank version no. AF184971.1 GI:6013324

Genbank record update date: Mar. 10, 2010 10:00 PM

Polypeptide

Genbank accession no. AAF01320

Genbank version no. AAF01320.1 GI:6013325

Genbank record update date: Mar. 10, 2010 10:00 PM

Cross References

Clark H.F., et al Genome Res. 13, 2265-2270, 2003; Mungall A. J., et alNature 425, 805-811, 2003; Blumberg H., et al Cell 104, 9-19, 2001;Dumoutier L., et al J. Immunol. 167, 3545-3549, 2001; Parrish-Novak J.,et al J. Biol. Chem. 277, 47517-47523, 2002; Pletnev S., et al (2003)Biochemistry 42:12617-12624; Sheikh F., et al (2004) J. Immuna 172,2006-2010; EP1394274 (Example 11); US2004/005320 (Example 5);WO2003/029262 (Page 74-75); WO2003/002717 (claim 2; Page 63);WO2002/22153 (Page 45-47); US2002/042366 (Page 20-21); WO2001/46261(Page 57-59); WO2001/46232 (Page 63-65); WO98/37193 (claim 1; Page55-59); Accession: Q9UHF4; Q6UWA9; Q96SH8; EMBL; AF184971; AAF01320.1.

(21) Brevican (BCAN, BEHAB)

Nucleotide

Genbank accession no AF229053

Genbank version no. AF229053.1 GI:10798902

Genbank record update date: Mar. 11, 2010 12:58 AM

Polypeptide

Genbank accession no. AAG23135

Genbank version no. AAG23135.1 GI:10798903

Genbank record update date: Mar. 11, 2010 12:58 AM

Cross References

Gary S. C., et al Gene 256, 139-147, 2000; Clark H.F., et al Genome Res.13, 2265-2270, 2003; Strausberg R. L., et al Proc. Natl. Acad. Sci.U.S.A. 99, 16899-16903, 2002; 52); US2003/186372 (claim 11);US2003/186373 (claim 11); US2003/119131 (claim 1; FIG. 52);US2003/119122 (claim 1; FIG. 52); US2003/119126 (claim 1); US2003/119121(claim 1; FIG. 52); US2003/119129 (claim 1); US2003/119130 (claim 1);US2003/119128 (claim 1; FIG. 52); US2003/119125 (claim 1); WO2003/016475(claim 1); WO2002/02634 (claim 1)

(22) EphB2R (DRT, ERK, Hek5, EPHT3, Tyro5)

Nucleotide

Genbank accession no NM_004442

Genbank version no. NM_004442.6 GI:111118979

Genbank record update date: Sep. 8, 2012 04:43 PM

Polypeptide

Genbank accession no. NP_004433

Genbank version no. NP_004433.2 GI:21396504

Genbank record update date: Sep. 8, 2012 04:43 PM

Cross References

Chan, J. and Watt, V. M., Oncogene 6 (6), 1057-1061 (1991) Oncogene 10(5):897-905 (1995), Annu. Rev. Neurosci. 21:309-345 (1998), Int. Rev.Cytol. 196:177-244 (2000)); WO2003042661 (claim 12); WO200053216 (claim1; Page 41); WO2004065576 (claim 1); WO2004020583 (claim 9);WO2003004529 (Page 128-132); WO200053216 (claim 1; Page 42); MIM:600997.

(23) ASLG659 (B7h)

Nucleotide

Genbank accession no. AX092328

Genbank version no. AX092328.1 GI:13444478

Genbank record update date: Jan. 26, 2011 07:37 AM

Cross References

US2004/0101899 (claim 2); WO2003104399 (claim 11); WO2004000221 (FIG.3); US2003/165504 (claim 1); US2003/124140 (Example 2); US2003/065143(FIG. 60); WO2002/102235 (claim 13; Page 299); US2003/091580 (Example2); WO2002/10187 (claim 6; FIG. 10); WO2001/94641 (claim 12; FIG. 7b);WO2002/02624 (claim 13; FIG. 1A-1B); US2002/034749 (claim 54; Page45-46); WO2002/06317 (Example 2; Page 320-321, claim 34; Page 321-322);WO2002/71928 (Page 468-469); WO2002/02587 (Example 1; FIG. 1);WO2001/40269 (Example 3; Pages 190-192); WO2000/36107 (Example 2; Page205-207); WO2004/053079 (claim 12); WO2003/004989 (claim 1);WO2002/71928 (Page 233-234, 452-453); WO 01/16318.

(24) PSCA (Prostate stem cell antigen precursor)

Nucleotide

Genbank accession no AJ297436

Genbank version no. AJ297436.1 GI:9367211

Genbank record update date: Feb. 1, 2011 11:25 AM

Polypeptide

Genbank accession no. CAB97347

Genbank version no. CAB97347.1 GI:9367212

Genbank record update date: Feb. 1, 2011 11:25 AM

Cross References

Reiter R. E., et al Proc. Natl. Acad. Sci. U.S.A. 95, 1735-1740, 1998;Gu Z., et al Oncogene 19, 1288-1296, 2000; Biochem. Biophys. Res.Commun. (2000) 275(3):783-788; WO2004/022709; EP1394274 (Example 11);US2004/018553 (claim 17); WO2003/008537 (claim 1); WO2002/81646 (claim1; Page 164); WO2003/003906 (claim 10; Page 288); WO2001/40309 (Example1; FIG. 17); US2001/055751 (Example 1; FIG. 1b); WO2000/32752 (claim 18;FIG. 1); WO98/51805 (claim 17; Page 97); WO98/51824 (claim 10; Page 94);WO98/40403 (claim 2; FIG. 1B); Accession: 043653; EMBL; AF043498;AAC39607.1

(25) GEDA

Nucleotide

Genbank accession no AY260763

Genbank version no. AY260763.1 GI:30102448

Genbank record update date: Mar. 11, 2010 02:24 AM

Polypeptide

Genbank accession no. AAP14954

Genbank version no. AAP14954.1 GI:30102449

Genbank record update date: Mar. 11, 2010 02:24 AM

Cross References

AP14954 lipoma HMGIC fusion-partnerlike protein/pid=AAP14954.1—Homosapiens (human); WO2003/054152 (claim 20); WO2003/000842 (claim 1);WO2003/023013 (Example 3, claim 20); US2003/194704 (claim 45);GI:30102449;

(26) BAFF-R (B cell -activating factor receptor, BLyS receptor 3, BR3)

Nucleotide

Genbank accession no AF116456

Genbank version no. AF116456.1 GI:4585274

Genbank record update date: Mar. 10, 2010 09:44 PM

Polypeptide

Genbank accession no. AAD25356

Genbank version no. AAD25356.1 GI:4585275

Genbank record update date: Mar. 10, 2010 09:44 PM

Cross References

BAFF receptor/pid=NP_443177.1—Homo sapiens: Thompson, J. S., et alScience 293 (5537), 2108-2111 (2001); WO2004/058309; WO2004/011611;WO2003/045422 (Example; Page 32-33); WO2003/014294 (claim 35; FIG. 6B);WO2003/035846 (claim 70; Page 615-616); WO2002/94852 (Col 136-137);WO2002/38766 (claim 3; Page 133); WO2002/24909 (Example 3; FIG. 3);MIM:606269; NP_443177.1; NM_052945_1; AF132600

(27) CD22 (B-cell receptor CD22-Bisoform, BL-CAM, Lyb-8, Lyb8, SIGLEC-2,FLJ22814)

Nucleotide

Genbank accession no AK026467

Genbank version no. AK026467.1 GI:10439337

Genbank record update date: Sep. 11, 2006 11:24 PM

Polypeptide

Genbank accession no. BAB15489

Genbank version no. BAB15489.1 GI:10439338

Genbank record update date: Sep. 11, 2006 11:24 PM

Cross References

Wilson et al (1991) J. Exp. Med. 173:137-146; WO2003/072036 (claim 1;FIG. 1); IM:107266; NP_001762.1; NM_001771_1.

(27a) CD22 (CD22 molecule)

Nucleotide

Genbank accession no X52785

Genbank version no. X52785.1 GI:29778

Genbank record update date: Feb. 2, 2011 10:09 AM

Polypeptide

Genbank accession no. CAA36988

Genbank version no. CAA36988.1 GI:29779

Genbank record update date: Feb. 2, 2011 10:09 AM

Cross References

Stamenkovic I. et al., Nature 345 (6270), 74-77 (1990)

Other Information

Official Symbol: CD22

Other Aliases: SIGLEC-2, SIGLEC2

Other Designations: B-cell receptor CD22; B-lymphocyte cell adhesionmolecule; BL-CAM; CD22 antigen; T-cell surface antigen Leu-14; sialicacid binding Ig-like lectin 2; sialic acid-binding Ig-like lectin 2

Antibodies

G5/44 (Inotuzumab): DiJoseph J F., et al Cancer Immunol Immunother. 2005January; 54(1):11-24.

Epratuzumab-Goldenberg D M., et al Expert Rev Anticancer Ther. 6(10):1341-53, 2006.

(28) CD79a (CD79A, CD79alpha), immunoglobulin-associated alpha, a Bcell-specific protein that covalently interacts with Ig beta (CD79B) andforms a complex on the surface with Ig M molecules, transduces a signalinvolved in B-cell differentiation), pl: 4.84, MW: 25028 TM: 2

[P] Gene Chromosome: 19q13.2).

Nucleotide

Genbank accession no NM_001783

Genbank version no. NM_001783.3 GI:90193587

Genbank record update date: Jun. 26, 2012 01:48 PM

Polypeptide

Genbank accession no. NP_001774

Genbank version no. NP_001774.1 GI:4502685

Genbank record update date: Jun. 26, 2012 01:48 PM

Cross References

WO2003/088808, US2003/0228319; WO2003/062401 (claim 9); US2002/150573(claim 4, pages 13-14); WO99/58658 (claim 13, FIG. 16); WO92/07574 (FIG.1); U.S. Pat. No. 5,644,033; Ha et al (1992) J. Immunol.148(5):1526-1531; Muller et al (1992) Eur. J. Immunol. 22:1621-1625;Hashimoto et al (1994) Immunogenetics 40(4):287-295; Preud'homme et al(1992) Clin. Exp. Immunol. 90(1):141-146; Yu et al (1992) J. Immunol.148(2) 633-637; Sakaguchi et al (1988) EMBO J. 7(11):3457-3464

(29) CXCR5 (Burkitt's lymphoma receptor 1, a G protein-coupled receptorthat is activated by the CXCL13 chemokine, functions in lymphocytemigration and humoral defense, plays a role in HIV-2 infection andperhaps development of AIDS, lymphoma, myeloma, and leukemia); 372 aa,pl: 8.54 MW: 41959 TM: 7 [P] Gene Chromosome: 11q23.3,

Nucleotide

Genbank accession no NM_001716

Genbank version no. NM_001716.4 GI:342307092

Genbank record update date: Sep. 30, 2012 01:49 PM

Polypeptide

Genbank accession no. NP_001707

Genbank version no. NP_001707.1 GI:4502415

Genbank record update date: Sep. 30, 2012 01:49 PM

Cross References

WO2004/040000; WO2004/015426; US2003/105292 (Example 2); U.S. Pat. No.6,555,339 (Example 2); WO2002/61087 (FIG. 1); WO2001/57188 (claim 20,page 269); WO2001/72830 (pages 12-13); WO2000/22129 (Example 1, pages152-153, Example 2, pages 254-256); WO99/28468 (claim 1, page 38); U.S.Pat. No. 5,440,021 (Example 2, col 49-52); WO94/28931 (pages 56-58);WO92/17497 (claim 7, FIG. 5); Dobner et al (1992) Eur. J. Immunol.22:2795-2799; Barella et al (1995) Biochem. J. 309:773-779

(30) HLA-DOB (Beta subunit of MHC class II molecule (la antigen) thatbinds peptides and presents them to CD4+T lymphocytes); 273 aa, pl:6.56, MW: 30820. TM: 1 [P]Gene Chromosome: 6p21.3)

Nucleotide

Genbank accession no NM_002120

Genbank version no. NM_002120.3 GI:118402587

Genbank record update date: Sep. 8, 2012 04:46 PM

Polypeptide

Genbank accession no. NP_002111

Genbank version no. NP_002111.1 GI:4504403

Genbank record update date: Sep. 8, 2012 04:46 PM

Cross References

Tonnelle et al (1985) EMBO J. 4(11):2839-2847; Jonsson et al (1989)Immunogenetics 29(6):411-413; Beck et al (1992) J. Mol. Biol.228:433-441; Strausberg et al (2002) Proc. Natl. Acad. Sci USA99:16899-16903; Servenius et al (1987) J. Biol. Chem. 262:8759-8766;Beck et al (1996) J. Mol. Biol. 255:1-13; Naruse et al (2002) TissueAntigens 59:512-519; WO99/58658 (claim 13, FIG. 15); U.S. Pat. No.6,153,408 (Col 35-38); U.S. Pat. No. 5,976,551 (col 168-170); U.S. Pat.No. 6,011,146 (col 145-146); Kasahara et al (1989) Immunogenetics30(1):66-68; Larhammar et al (1985) J. Biol. Chem. 260(26):14111-14119

(31) P2X5 (Purinergic receptor P2X ligand-gated ion channel 5, an ionchannel gated by extracellular ATP, may be involved in synaptictransmission and neurogenesis, deficiency may contribute to thepathophysiology of idiopathic detrusor instability); 422 aa), pl: 7.63,MW: 47206 TM: 1 [P] Gene Chromosome: 17p13.3).

Nucleotide

Genbank accession no NM_002561

Genbank version no. NM_002561.3 GI:325197202

Genbank record update date: Jun. 27, 2012 12:41 AM

Polypeptide

Genbank accession no. NP_002552

Genbank version no. NP_002552.2 GI:28416933

Genbank record update date: Jun. 27, 2012 12:41 AM

Cross References

Le et al (1997) FEBS Lett. 418(1-2):195-199; WO2004/047749;WO2003/072035 (claim 10); Touchman et al (2000) Genome Res. 10:165-173;WO2002/22660 (claim 20); WO2003/093444 (claim 1); WO2003/087768 (claim1); WO2003/029277 (page 82) (32) CD72 (B-cell differentiation antigenCD72, Lyb-2); 359 aa, pl: 8.66, MW: 40225, TM: 1 5 [P] Gene Chromosome:9p13.3).

Nucleotide

Genbank accession no NM_001782

Genbank version no. NM_001782.2 GI:194018444

Genbank record update date: Jun. 26, 2012 01:43 PM

Polypeptide

Genbank accession no. NP_001773

Genbank version no. NP_001773.1 GI:4502683

Genbank record update date: Jun. 26, 2012 01:43 PM

Cross References

WO2004042346 (claim 65); WO2003/026493 (pages 51-52, 57-58);WO2000/75655 (pages 105-106); Von Hoegen et al (1990) J. Immunol.144(12):4870-4877; Strausberg et al (2002) Proc. Natl. Acad. Sci USA99:16899-16903.

(33) LY64 (Lymphocyte antigen 64 (RP105), type I membrane protein of theleucine rich repeat (LRR) family, regulates B-cell activation andapoptosis, loss of function is associated

with increased disease activity in patients with systemic lupuserythematosis); 661 aa, pl: 6.20, MW: 74147 TM: 1 [P] Gene Chromosome:5q12).

Nucleotide

Genbank accession no NM_005582

Genbank version no. NM_005582.2 GI:167555126

Genbank record update date: Sep. 2, 2012 01:50 PM

Polypeptide

Genbank accession no. NP_005573

Genbank version no. NP_005573.2 GI:167555127

Genbank record update date: Sep. 2, 2012 01:50 PM

Cross References

US2002/193567; WO97/07198 (claim 11, pages 39-42); Miura et al (1996)Genomics 38(3):299-304; Miura et al (1998) Blood 92:2815-2822;WO2003/083047; WO97/44452 (claim 8, pages 57-61); WO2000/12130 (pages24-26).

(34) FcRH1 (Fc receptor-like protein 1, a putative receptor for theimmunoglobulin Fc domain that contains C2 type Ig-like and ITAM domains,may have a role in B-lymphocyte differentiation); 429 aa, pl: 5.28, MW:46925 TM: 1 [P] Gene Chromosome: 1q21-1q22)

Nucleotide

Genbank accession no NM_052938

Genbank version no. NM_052938.4 GI:226958543

Genbank record update date: Sep. 2, 2012 01:43 PM

Polypeptide

Genbank accession no. NP_443170

Genbank version no. NP_443170.1 GI:16418419

Genbank record update date: Sep. 2, 2012 01:43 PM

Cross References

WO2003/077836; WO2001/38490 (claim 6, FIG. 18E-1-18-E-2); Davis et al(2001) Proc. Natl. Acad. Sci USA 98(17):9772-9777; WO2003/089624 (claim8); EP1347046 (claim 1); WO2003/089624 (claim 7).

(35) IRTA2 (Immunoglobulin superfamily receptor translocation associated2, a putative immunoreceptor with possible roles in B cell developmentand lymphoma genesis; deregulation of the gene by translocation occursin some B cell malignancies); 977 aa, pl: 6.88, MW: 106468, TM: 1 [P]Gene Chromosome: 1q21)

Nucleotide

Genbank accession no AF343662

Genbank version no. AF343662.1 GI:13591709

Genbank record update date: Mar. 11, 2010 01:16 AM

Polypeptide

Genbank accession no. AAK31325

Genbank version no. AAK31325.1 GI:13591710

Genbank record update date: Mar. 11, 2010 01:16 AM

Cross References

AF343663, AF343664, AF343665, AF369794, AF397453, AK090423, AK090475,AL834187, AY358085; Mouse:AK089756, AY158090, AY506558; NP_112571.1;WO2003/024392 (claim 2, FIG. 97); Nakayama et al (2000) Biochem.Biophys. Res. Commun. 277(1):124-127; WO2003/077836; WO2001/38490 (claim3, FIG. 18B-1-18B-2).

(36) TENB2 (TMEFF2, tomoregulin, TPEF, HPP1, TR, putative transmembraneproteoglycan, related to the EGF/heregulin family of growth factors andfollistatin); 374 aa)

Nucleotide

Genbank accession no AF179274

Genbank version no. AF179274.2 GI:12280939

Genbank record update date: Mar. 11, 2010 01:05 AM

Polypeptide

Genbank accession no. AAD55776

Genbank version no. AAD55776.2 GI:12280940

Genbank record update date: Mar. 11, 2010 01:05 AM

Cross References

NCBI Accession: AAD55776, AAF91397, AAG49451, NCBI RefSeq: NP_057276;NCBI Gene: 23671; OMIM: 605734; SwissProt Q9UIK5; AY358907, CAF85723,CQ782436; WO2004/074320; JP2004113151; WO2003/042661; WO2003/009814;EP1295944 (pages 69-70); WO2002/30268 (page 329); WO2001/90304;US2004/249130; US2004/022727; WO2004/063355; US2004/197325;US2003/232350; US2004/005563; US2003/124579; Horie et al (2000) Genomics67:146-152; Uchida et al (1999) Biochem. Biophys. Res. Commun.266:593-602; Liang et al (2000) Cancer Res. 60:4907-12; Glynne-Jones etal (2001) Int J Cancer. October 15; 94(2):178-84.

(37) PSMA-FOLH1 (Folate hydrolase (prostate-specific membrane antigen)1)

Nucleotide

Genbank accession no M99487

Genbank version no. M99487.1 GI:190663

Genbank record update date: Jun. 23, 2010 08:48 AM

Polypeptide

Genbank accession no. AAA60209

Genbank version no. AAA60209.1 GI:190664

Genbank record update date: Jun. 23, 2010 08:48 AM

Cross References

Israeli R. S., et al Cancer Res. 53 (2), 227-230 (1993)

Other Information

Official Symbol: FOLH1

Other Aliases: GIG27, FGCP, FOLH, GCP2, GCPII, NAALAD1, NAALAdase, PSM,PSMA, mGCP

Other Designations: N-acetylated alpha-linked acidic dipeptidase 1;N-acetylated-alpha-linked acidic dipeptidase I; NAALADase I; cellgrowth-inhibiting gene 27 protein; folylpoly-gamma-glutamatecarboxypeptidase; glutamate carboxylase II; glutamate carboxypeptidase2; glutamate carboxypeptidase II; membrane glutamate carboxypeptidase;prostate specific membrane antigen variant F;pteroylpoly-gamma-glutamate carboxypeptidase

Antibodies

U.S. Pat. No. 7,666,425:

Antibodies produces by Hybridomas having the following ATCCreferences:ATCC accession No. HB-12101, ATCC accession No. HB-12109,ATCC accession No. HB-12127 and ATCC accession No. HB-12126.

Proscan: a monoclonal antibody selected from the group consisting of8H12, 3E11, 17G1, 29B4, 30C1 and 20F2 (U.S. Pat. No. 7,811,564; MoffettS., et al Hybridoma (Larchmt). 2007 December; 26(6):363-72).

Cytogen: monoclonal antibodies 7E11-05 (ATCC accession No. HB 10494) and9H10-A4 (ATCC accession No. HB11430)—U.S. Pat. No. 5,763,202

GlycoMimetics: NUH2—ATCC accession No. HB 9762 (U.S. Pat. No. 7,135,301)

Human Genome Science: HPRAJ70—ATCC accession No. 97131 (U.S. Pat. No.6,824,993); Amino acid sequence encoded by the cDNA clone (HPRAJ70)deposited as American Type Culture Collection (“ATCC”) Deposit No. 97131

Medarex: Anti-PSMA antibodies that lack fucosyl residues—U.S. Pat. No.7,875,278

Mouse anti-PSMA antibodies include the 3F5.4G6, 3D7.1.1, 4E10-1.14,3E11, 4D8, 3E6, 3C9, 2C7, 1G3, 3C4, 3C6, 4D4, 1G9, 5C8B9, 3G6, 4C8B9,and monoclonal antibodies. Hybridomas secreting 3F5.4G6, 3D7.1.1,4E10-1.14, 3E11, 4D8, 3E6, 3C9, 2C7, 1G3, 3C4, 3C6, 4D4, 1G9, 5C8B9, 3G6or 4C8B9 have been publicly deposited and are described in U.S. Pat. No.6,159,508. Relevant hybridomas have been publicly deposited and aredescribed in U.S. Pat. No. 6,107,090. Moreover, humanized anti-PSMAantibodies, including a humanized version of J591, are described infurther detail in PCT Publication WO 02/098897.

Other mouse anti-human PSMA antibodies have been described in the art,such as mAb 107-1A4 (Wang, S. et al. (2001) Int. J. Cancer 92:871-876)and mAb 2C9 (Kato, K. et al. (2003) Int. J. Urol. 10:439-444).

Examples of human anti-PSMA monoclonal antibodies include the 4A3, 7F12,8C12, 8A11, 16F9, 2A10, 2C6, 2F5 and 1C3 antibodies, isolated andstructurally characterized as originally described in PCT PublicationsWO 01/09192 and WO 03/064606 and in U.S. Provisional Application Ser.No. 60/654,125, entitled “Human Monoclonal Antibodies to ProstateSpecific Membrane Antigen (PSMA)”, filed on Feb. 18, 2005. The V.sub.Hamino acid sequences of 4A3, 7F12, 8C12, 8A11, 16F9, 2A10, 2C6, 2F5 and1C3 are shown in SEQ ID NOs: 1-9, respectively. The V.sub.L amino acidsequences of 4A3, 7F12, 8C12, 8A11, 16F9, 2A10, 2C6, 2F5 and 1C3 areshown in SEQ ID NOs: 10-18, respectively.

Other human anti-PSMA antibodies include the antibodies disclosed in PCTPublication WO 03/034903 and US Application No. 2004/0033229.

NW Biotherapeutics: A hybridoma cell line selected from the groupconsisting of 3F5.4G6 having ATCC accession number HB12060, 3D7-1.I.having ATCC accession number HB12309, 4E10-1.14 having ATCC accessionnumber HB12310, 3E11 (ATCC HB12488), 4D8 (ATCC HB12487), 3E6 (ATCCHB12486), 3C9 (ATCC HB12484), 2C7 (ATCC HB12490), 1G3 (ATCC HB12489),3C4 (ATCC HB12494), 3C6 (ATCC HB12491), 4D4 (ATCC HB12493), 1G9 (ATCCHB12495), 5C8B9 (ATCC HB12492) and 3G6 (ATCC HB12485)—see U.S. Pat. No.6,150,508

PSMA Development Company/Progenics/Cytogen—Seattle Genetics: mAb 3.9,produced by the hybridoma deposited under ATCC Accession No. PTA-3258 ormAb 10.3, produced by the hybridoma deposited under ATCC Accession No.PTA-3347—U.S. Pat. No. 7,850,971

PSMA Development Company-Compositions of PSMA antibodies (US20080286284, Table 1)

-   -   This application is a divisional of U.S. patent application Ser.        No. 10/395,894, filed on Mar. 21, 2003 (U.S. Pat. No. 7,850,971)

University Hospital Freiburg, Germany—mAbs 3/A12, 3/E7, and 3/F11 (WolfP., et al Prostate. 2010 Apr. 1; 70(5):562-9).

(38) SST (Somatostatin Receptor; note that there are 5 subtypes)

(38.1) SSTR2 (Somatostatin receptor 2)

Nucleotide

Genbank accession no NM_001050

Genbank version no. NM_001050.2 GI:44890054

Genbank record update date: Aug. 19, 2012 01:37 PM

Polypeptide

Genbank accession no. NP_001041

Genbank version no. NP_001041.1 GI:4557859

Genbank record update date: Aug. 19, 2012 01:37 PM

Cross References

Yamada Y., et al Proc. Natl. Acad. Sci. U.S.A. 89 (1), 251-255 (1992);Susini C., et al Ann Oncol. 2006 December; 17(12):1733-42

Other Information

Official Symbol: SSTR2

Other Designations: SRIF-1; SS2R; somatostatin receptor type 2

(38.2) SSTR5 (Somatostatin receptor 5)

Nucleotide

Genbank accession no D16827

Genbank version no. D16827.1 GI:487683

Genbank record update date: Aug. 1, 2006 12:45 PM

Polypeptide

Genbank accession no. BAA04107

Genbank version no. BAA04107.1 GI:487684

Genbank record update date: Aug. 1, 2006 12:45 PM

Cross References

Yamada, Y., et al Biochem. Biophys. Res. Commun. 195 (2), 844-852 (1993)

Other Information

Official Symbol: SSTR5

Other Aliases: SS-5-R

Other Designations: Somatostatin receptor subtype 5; somatostatinreceptor type 5

(38.3) SSTR1

(38.4)SSTR3

(38.5) SSTR4

AvB6—Both subunits (39+40)

(39) ITGAV (Integrin, alpha V;

Nucleotide

Genbank accession no M14648 J02826 M18365

Genbank version no. M14648.1 GI:340306

Genbank record update date: Jun. 23, 2010 08:56 AM

Polypeptide

Genbank accession no. AAA36808

Genbank version no. AAA36808.1 GI:340307

Genbank record update date: Jun. 23, 2010 08:56 AM

Cross References

Suzuki S., et al Proc. Natl. Acad. Sci. U.S.A. 83 (22), 8614-8618 (1986)

Other Information

Official Symbol: ITGAV

Other Aliases: CD51, MSK8, VNRA, VTNR

Other Designations: antigen identified by monoclonal antibody L230;integrin alpha-V; integrin alphaVbeta3; integrin, alpha V (vitronectinreceptor, alpha polypeptide, antigen CD51); vitronectin receptor subunitalpha

(40) ITGB6 (Integrin, beta 6)

Nucleotide

Genbank accession no NM_000888

Genbank version no. NM_000888.3 GI:9966771

Genbank record update date: Jun. 27, 2012 12:46 AM

Polypeptide

Genbank accession no. NP_000879

Genbank version no. NP_000879.2 GI:9625002

Genbank record update date: Jun. 27, 2012 12:46 AM

Cross References

Sheppard D. J., et al Biol. Chem. 265 (20), 11502-11507 (1990)

Other Information

Official Symbol: ITGB6

Other Designations: integrin beta-6

Antibodies

Biogen: U.S. Pat. No. 7,943,742—Hybridoma clones 6.3G9 and 6.8G6 weredeposited with the ATCC, accession numbers ATCC PTA-3649 and -3645,respectively.

Biogen: U.S. Pat. No. 7,465,449—In some embodiments, the antibodycomprises the same heavy and light chain polypeptide sequences as anantibody produced by hybridoma 6.1A8, 6.3G9, 6.8G6, 6.2B1, 6.2B10,6.2A1, 6.2E5, 7.1G10, 7.7G5, or 7.105.

Centocor (J&J): U.S. Pat. Nos. 7,550,142; 7,163,681

-   -   For example in U.S. Pat. No. 7,550,142—an antibody having human        heavy chain and human light chain variable regions comprising        the amino acid sequences shown in SEQ ID NO: 7 and SEQ ID NO: 8.

Seattle Genetics: 15H3 (Ryan M C., et al Cancer Res Apr. 15, 2012; 72(8Supplement): 4630)

(41) CEACAM5 (Carcinoembryonic antigen-related cell adhesion molecule 5)

Nucleotide

Genbank accession no M17303

Genbank version no. M17303.1 GI:178676

Genbank record update date: Jun. 23, 2010 08:47 AM

Polypeptide

Genbank accession no. AAB59513

Genbank version no. AAB59513.1 GI:178677

Genbank record update date: Jun. 23, 2010 08:47 AM

Cross References

Beauchemin N., et al Mol. Cell. Biol. 7 (9), 3221-3230 (1987)

Other Information

Official Symbol: CEACAM5

Other Aliases: CD66e, CEA

Other Designations: meconium antigen 100

Antibodies

AstraZeneca-Medlmmune:US 20100330103; US20080057063;

US20020142359

-   -   for example an antibody having complementarity determining        regions (CDRs) with the following sequences: heavy chain;        CDR1—DNYMH, CDR2—WIDPENGDTE YAPKFRG, CDR3—LIYAGYLAMD Y; and        light chain CDR1—SASSSVTYMH, CDR2—STSNLAS, CDR3—QQRSTYPLT.    -   Hybridoma 806.077 deposited as European Collection of Cell        Cultures (ECACC) deposit no. 96022936.

Research Corporation Technologies, Inc.: U.S. Pat. No. 5,047,507

Bayer Corporation: U.S. Pat. No. 6,013,772

BioAlliance: U.S. Pat. No. 7,982,017; 7,674,605

-   -   U.S. Pat. No. 7,674,605        -   an antibody comprising the heavy chain variable region            sequence from the amino acid sequence of SEQ ID NO: 1, and            the light chain variable region sequence from the amino acid            sequence of SEQ ID NO:2.        -   an antibody comprising the heavy chain variable region            sequence from the amino acid sequence of SEQ ID NO:5, and            the light chain variable region sequence from the amino acid            sequence of SEQ ID NO:6.

Celltech Therapeutics Limited: U.S. Pat. No. 5,877,293

The Dow Chemical Company: U.S. Pat. Nos. 5,472,693; 6,417,337; 6,333,405

-   -   U.S. Pat. No. 5,472,693—for example, ATCC No. CRL-11215    -   U.S. Pat. No. 6,417,337—for example, ATCC CRL-12208    -   U.S. Pat. No. 6,333,405—for example, ATCC CRL-12208

Immunomedics, Inc: U.S. Pat. Nos. 7,534,431; 7,230,084; 7,300,644;6,730,300;

-   -   US20110189085        -   an antibody having CDRs of the light chain variable region            comprise: CDR1 comprises KASQDVGTSVA (SEQ ID NO: 20); CDR2            comprises WTSTRHT (SEQ ID NO: 21); and CDR3 comprises            QQYSLYRS (SEQ ID NO: 22);        -   and the CDRs of the heavy chain variable region of said            anti-CEA antibody comprise: CDR1 comprises TYWMS (SEQ ID NO:            23); CDR2 comprises EIHPDSSTINYAPSLKD (SEQ ID NO: 24); and            CDR3 comprises LYFGFPWFAY (SEQ ID NO: 25).    -   US20100221175; US20090092598; US20070202044; US20110064653;        US20090185974; US20080069775.

(42) MET (met proto-oncogene; hepatocyte growth factor receptor)

Nucleotide

Genbank accession no M35073

Genbank version no. M35073.1 GI:187553

Genbank record update date: Mar. 6, 2012 11:12 AM

Polypeptide

Genbank accession no. AAA59589

Genbank version no. AAA59589.1 GI:553531

Genbank record update date: Mar. 6, 2012 11:12 AM

Cross References

Dean M., et al Nature 318 (6044), 385-388 (1985)

Other Information

Official Symbol: MET

Other Aliases: AUTS9, HGFR, RCCP2, c-Met

Other Designations: HGF receptor; HGF/SF receptor; SF receptor;hepatocyte growth factor receptor; met proto-oncogene tyrosine kinase;proto-oncogene c-Met; scatter factor receptor; tyrosine-protein kinaseMet

Antibodies

Abgenix/Pfizer: US20100040629

-   -   for example, the antibody produced by hybridoma 13.3.2 having        American Type Culture Collection (ATCC) accession number        PTA-5026; the antibody produced by hybridoma 9.1.2 having ATCC        accession number PTA-5027; the antibody produced by hybridoma        8.70.2 having ATCC accession number PTA-5028; or the antibody        produced by hybridoma 6.90.3 having ATCC accession number        PTA-5029.

Amgen/Pfizer: US20050054019

-   -   for example, an antibody comprising a heavy chain having the        amino acid sequences set forth in SEQ ID NO: 2 where X2 is        glutamate and X4 is serine and a light chain having the amino        acid sequence set forth in SEQ ID NO: 4 where X8 is alanine,        without the signal sequences; an antibody comprising a heavy        chain having the amino acid sequences set forth in SEQ ID NO: 6        and a light chain having the amino acid sequence set forth in        SEQ ID NO: 8, without the signal sequences; an antibody        comprising a heavy chain having the amino acid sequences set        forth in SEQ ID NO: 10 and a light chain having the amino acid        sequence set forth in SEQ ID NO: 12, without the signal        sequences; or an antibody comprising a heavy chain having the        amino acid sequences set forth in SEQ ID NO: 14 and a light        chain having the amino acid sequence set forth in SEQ ID NO: 16,        without the signal sequences.

Agouron Pharmaceuticals (Now Pfizer): US20060035907

Eli Lilly: US20100129369

Genentech: U.S. Pat. No. 5,686,292; US20100028337; US20100016241;US20070129301; US20070098707; US20070092520, US20060270594;US20060134104; US20060035278; US20050233960; US20050037431

-   -   U.S. Pat. No. 5,686,292—for example, ATCC HB-11894 and ATCC        HB-11895    -   US 20100016241—for example, ATCC HB-11894 (hybridoma 1A3.3.13)        or HB-11895 (hybridoma 5D5.11.6)

National Defense Medical Center, Taiwan: Lu R M., et al Biomaterials.2011 April; 32(12):3265-74.

Novartis: US20090175860

-   -   for example, an antibody comprising the sequences of CDR1, CDR2        and CDR3 of heavy chain 4687, wherein the sequences of CDR1,        CDR2, and CDR3 of heavy chain 4687 are residues 26-35, 50-65,        and 98-102, respectively, of SEQ ID NO: 58; and the sequences of        CDR1, CDR2, and CDR3 of light chain 5097, wherein the sequences        of CDR1, CDR2, and CDR3 of light chain 5097 are residues 24-39,        55-61, and 94-100 of SEQ ID NO: 37.

Pharmacia Corporation: US20040166544

Pierre Fabre: US20110239316, US20110097262, US20100115639

Sumsung: US 20110129481—for example a monoclonal antibody produced froma hybridoma cell having accession number KCLRF-BP-00219 or accessionnumber of KCLRF-BP-00223.

Samsung: US 20110104176—for example an antibody produced by a hybridomacell having Accession Number: KCLRF-BP-00220.

University of Turin Medical School: DN-30 Pacchiana G., et al J BiolChem. 2010 Nov. 12; 285(46):36149-57

Van Andel Research Institute: Jiao Y., et al Mol Biotechnol. 2005September; 31(1):41-54.

(43) MUC1 (Mucin 1, cell surface associated)

Nucleotide

Genbank accession no J05581

Genbank version no. J05581.1 GI:188869

Genbank record update date: Jun. 23, 2010 08:48 AM

Polypeptide

Genbank accession no. AAA59876

Genbank version no. AAA59876.1 GI:188870

Genbank record update date: Jun. 23, 2010 08:48 AM

Cross References

Gendler S. J., et al J. Biol. Chem. 265 (25), 15286-15293 (1990)

Other Information

Official Symbol: MUC1

Other Aliases: RP11-263K19.2, CD227, EMA, H23AG, KL-6, MAM6, MUC-1,MUC-1/SEC, MUC-1/X, MUC1/ZD, PEM, PEMT, PUM

Other Designations: DF3 antigen; H23 antigen; breastcarcinoma-associated antigen DF3; carcinoma-associated mucin; episialin;krebs von den Lungen-6; mucin 1, transmembrane; mucin-1; peanut-reactiveurinary mucin; polymorphic epithelial mucin; tumor associated epithelialmucin; tumor-associated epithelial membrane antigen; tumor-associatedmucin

Antibodies

AltaRex—Quest Pharma Tech: U.S. Pat. No. 6,716,966—for example an Alt-1antibody produced by the hybridoma ATCC No PTA-975.

AltaRex—Quest Pharma Tech: U.S. Pat. No. 7,147,850

-   CRT: 5E5-Sørensen A L., et al Glycobiology vol. 16 no. 2 pp. 96-107,    2006; HMFG2-Burchell J., et al Cancer Res., 47, 5476-5482 (1987)

Glycotope GT-MAB: GT-MAB 2.5-GEX (Website:http://www.glycotope.com/pipeline/pankomab-gex)

Immunogen: U.S. Pat. No. 7,202,346

-   -   for example, antibody MJ-170: hybridoma cell line MJ-170 ATCC        accession no. PTA-5286Monoclonal antibody MJ-171: hybridoma cell        line MJ-171 ATCC accession no. PTA-5287; monoclonal antibody        MJ-172: hybridoma cell line MJ-172 ATCC accession no. PTA-5288;        or monoclonal antibody MJ-173: hybridoma cell line MJ-173 ATCC        accession no. PTA-5302

Immunomedics: U.S. Pat. No. 6,653,104

Ramot Tel Aviv Uni: U.S. Pat. No. 7,897,351

Regents Uni. CA: U.S. Pat. No. 7,183,388; US20040005647; US20030077676.

Roche GlycArt: U.S. Pat. No. 8,021,856

Russian National Cancer Research Center: Imuteran-Ivanov P K., et alBiotechnol J. 2007 July; 2(7):863-70

Technische Univ Braunschweig: (IIB6, HT186-B7, HT186-D11, HT186-G2,HT200-3A-C1, HT220-M-D1, HT220-M-G8)—Thie H., et al PLoS One. 2011 Jan.14; 6(1):e15921

(44) CA9 (Carbonic anhydrase IX)

Nucleotide

Genbank accession no. X66839

Genbank version no. X66839.1 GI:1000701

Genbank record update date: Feb. 2, 2011 10:15 AM

Polypeptide

Genbank accession no. CAA47315

Genbank version no. CAA47315.1 GI:1000702

Genbank record update date: Feb. 2, 2011 10:15 AM

Cross References

Pastorek J., et al Oncogene 9 (10), 2877-2888 (1994)

Other Information

Official Symbol: CA9

Other Aliases: CAIX, MN

Other Designations: CA-1X; P54/58N; RCC-associated antigen G250;RCC-associated protein G250; carbonate dehydratase IX; carbonicanhydrase 9; carbonic dehydratase; membrane antigen MN; pMW1; renal cellcarcinoma-associated antigen G250

Antibodies

Abgenix/Amgen: US20040018198

Affibody: Anti-CAIX Affibody molecules(http://www.affibody.com/en/Product-Portfolio/Pipeline/)

Bayer: U.S. Pat. No. 7,462,696

Bayer/Morphosys: 3ee9 mAb—Petrul H M., et al Mol Cancer Ther. 2012February; 11(2):340-9

Harvard Medical School: Antibodies G10, G36, G37, G39, G45, G57, G106,G119, G6, G27, G40 and G125. Xu C., et al PLoS One. 2010 Mar. 10;5(3):e9625

Institute of Virology, Slovak Academy of Sciences (Bayer)—U.S. Pat. No.5,955,075

-   -   for example, M75—ATCC Accession No. HB 11128 or MN12—ATCC        Accession No. HB 11647

Institute of Virology, Slovak Academy of Sciences: U.S. Pat. No.7,816,493

-   -   for example the M75 monoclonal antibody that is secreted from        the hybridoma VU-M75, which was deposited at the American Type        Culture Collection under ATCC No. HB 11128; or the V/10        monoclonal antibody secreted from the hybridoma V/10-VU, which        was deposited at the International Depository Authority of the        Belgian Coordinated Collection of Microorganisms (BCCM) at the        Laboratorium voor Moleculaire

Bioloqie-Plasmidencollectie (LMBP) at the Universeit Gent in Gent,Belgium, under Accession No. LMBP 6009CB.

Institute of Virology, Slovak Academy of Sciences US20080177046;US20080176310; US20080176258; US20050031623

Novartis: US20090252738

Wilex: U.S. Pat. No. 7,691,375—for example the antibody produced by thehybridoma cell line DSM ASC 2526.

Wilex: US20110123537; Rencarex: Kennett R H., et al Curr Opin Mol Ther.2003 February; 5(1):70-5

Xencor: US20090162382

(45) EGFRvIII (Epidermal growth factor receptor (EGFR), transcriptvariant 3,

Nucleotide

Genbank accession no. NM_201283

Genbank version no. NM_201283.1 GI:41327733

Genbank record update date: Sep. 30, 2012 01:47 PM

Polypeptide

Genbank accession no. NP_958440

Genbank version no. NP_958440.1 GI:41327734

Genbank record update date: Sep. 30, 2012 01:47 PM

Cross-References

Batra S K., et al Cell Growth Differ 1995; 6:1251-1259.

Antibodies:

U.S. Pat. Nos. 7,628,986 and 7,736,644 (Amgen)

-   -   For example, a heavy chain variable region amino acid sequence        selected from the group consisting of SEQ ID NO: 142 and        variants & a light chain variable region amino acid sequence        selected from the group consisting of: SEQ ID NO: 144 and        variants.

US20100111979 (Amgen)

For example, an antibody comprising a heavy chain amino acid sequencecomprising:

-   -   CDR1 consisting of a sequence selected from the group consisting        of the amino acid sequences for the CDR1 region of antibodies        13.1.2 (SEQ ID NO: 138), 131 (SEQ ID NO: 2), 170 (SEQ ID NO: 4),        150 (SEQ ID NO: 5), 095 (SEQ ID NO: 7), 250 (SEQ ID NO: 9), 139        (SEQ ID NO: 10), 211 (SEQ ID NO: 12), 124 (SEQ ID NO: 13), 318        (SEQ ID NO: 15), 342 (SEQ ID NO: 16), and 333 (SEQ ID NO: 17);        CDR2 consisting of a sequence selected from the group consisting        of the amino acid sequences for the CDR2 region of antibodies        13.1.2 (SEQ ID NO: 138), 131 (SEQ ID NO: 2), 170 (SEQ ID NO: 4),        150 (SEQ ID NO: 5), 095 (SEQ ID NO: 7), 250 (SEQ ID NO: 9), 139        (SEQ ID NO: 10), 211 (SEQ ID NO: 12), 124 (SEQ ID NO: 13), 318        (SEQ ID NO: 15), 342 (SEQ ID NO: 16), and 333 (SEQ ID NO: 17);        and    -   CDR3 consisting of a sequence selected from the group consisting        of the amino acid sequences for the CDR3 region of antibodies        13.1.2 (SEQ ID NO: 138), 131 (SEQ ID NO: 2), 170 (SEQ ID NO: 4),        150 (SEQ ID NO: 5), 095 (SEQ ID NO: 7), 250 (SEQ ID NO: 9), 139        (SEQ ID NO: 10), 211 (SEQ ID NO: 12), 124 (SEQ ID NO: 13), 318        (SEQ ID NO: 15), 342 (SEQ ID NO: 16), and 333 (SEQ ID NO: 17).

US20090240038 (Amgen)

-   -   For example, an antibody having at least one of the heavy or        light chain polypeptides comprises an amino acid sequence that        is at least 90% identical to the amino acid sequence selected        from the group consisting of: SEQ ID NO: 2, SEQ ID NO: 19, SEQ        ID NO: 142, SEQ ID NO: 144, and any combination thereof.

US20090175887 (Amgen)

-   -   For example, an antibody having a heavy chain amino acid        sequence selected from the group consisting of the heavy chain        amino acid sequence of antibody 13.1.2 (SEQ ID NO: 138), 131        (SEQ ID NO: 2), 170 (SEQ ID NO: 4), 150 (SEQ ID NO: 5), 095 (SEQ        ID NO: 7), 250 (SEQ ID NO: 9), 139 (SEQ ID NO: 10), 211 (SEQ ID        NO: 12), 124 (SEQ ID NO: 13), 318 (SEQ ID NO: 15), 342 (SEQ ID        NO: 16), and 333 (SEQ ID NO: 17).

US20090156790 (Amgen)

-   -   For example, antibody having heavy chain polypeptide and a light        chain polypeptide, wherein at least one of the heavy or light        chain polypeptides comprises an amino acid sequence that is at        least 90% identical to the amino acid sequence selected from the        group consisting of: SEQ ID NO: 2, SEQ ID NO: 19, SEQ ID NO: 12)        142, SEQ ID NO: 144, and any combination thereof.

US20090155282, US20050059087 and US20050053608 (Amgen)

-   -   For example, an antibody heavy chain amino acid sequence        selected from the group consisting of the heavy chain amino acid        sequence of antibody 13.1.2 (SEQ ID NO: 138), 131 (SEQ ID NO:        2), 170 (SEQ ID NO: 4), 150 (SEQ ID NO: 5), 095 (SEQ ID NO: 7),        250 (SEQ ID NO: 9), 139 (SEQ ID NO: 10), 211 (SEQ ID NO: 12),        124 (SEQ ID NO: 13), 318 (SEQ ID NO: 15), 342 (SEQ ID NO: 16),        and 333 (SEQ ID NO: 17).

MR1-1 (U.S. Pat. No. 7,129,332; Duke)

-   -   For example, a variant antibody having the sequence of SEQ ID        NO.18 with the substitutions S98P-T99Y in the CDR3 VH, and F92W        in CDR3 VL.

L8A4, H10, Y10 (Wikstrand C J., et al Cancer Res. 1995 Jul. 15;55(14):3140-8; Duke)

US20090311803 (Harvard University)

-   -   For example, SEQ ID NO:9 for antibody heavy chain variable        region, and SEQ ID NO: 3 for light chain variable region amino        acid sequences

US20070274991 (EMD72000, also known as matuzumab; Harvard University)

-   -   For example, SEQ ID NOs: 3 & 9 for light chain and heavy chain        respectively

U.S. Pat. No. 6,129,915 (Schering)

-   -   For example, SEQ. ID NOs: 1, 2, 3, 4, 5 and 6.

mAb CH12—Wang H., et al FASEB J. 2012 January; 26(1):73-80 (ShanghaiCancer Institute).

RAbDMvIII—Gupta P., et al BMC Biotechnol. 2010 Oct. 7; 10:72 (StanfordUniversity Medical Center).

mAb Ua30—Ohman L., et al Tumour Biol. 2002 March-April; 23(2):61-9(Uppsala University).

Han D G., et al Nan Fang Yi Ke Da Xue Xue Bao. 2010 January; 30(1):25-9(Xi'an Jiaotong University).

(46) CD33 (CD33 molecule)

Nucleotide

Genbank accession no. M_23197

Genbank version no. NM_23197.1 GI:180097

Genbank record update date: Jun. 23, 2010 08:47 AM

Polypeptide

Genbank accession no. AAA51948

Genbank version no. AAA51948.1 GI:188098

Genbank record update date: Jun. 23, 2010 08:47 AM

Cross-References

Simmons D., et al J. Immunol. 141 (8), 2797-2800 (1988)

Other Information

Official Symbol: CD33

Other Aliases: SIGLEC-3, SIGLEC3, p67

Other Designations: CD33 antigen (gp67); gp67; myeloid cell surfaceantigen CD33; sialic acid binding Ig-like lectin 3; sialic acid-bindingIg-like lectin

Antibodies

H195 (Lintuzumab)—Raza A., et al Leuk Lymphoma. 2009 August;50(8):1336-44; U.S. Pat. No. 6,759,045 (Seattle Genetics/Immunomedics)

mAb OKT9: Sutherland, D. R. et al. Proc Natl Acad Sci USA 78(7):4515-4519 1981, Schneider, C., et al J Biol Chem 257, 8516-8522 (1982)

mAb E6: Hoogenboom, H. R., et al J Immunol 144, 3211-3217 (1990)

U.S. Pat. No. 6,590,088 (Human Genome Sciences)

-   -   For example, SEQ ID NOs: 1 and 2 and ATCC accession no. 97521

U.S. Pat. No. 7,557,189 (Immunogen)

-   -   For example, an antibody or fragment thereof comprising a heavy        chain variable region which comprises three CDRs having the        amino acid sequences of SEQ ID NOs:1-3 and a light chain        variable region comprising three CDRs having the amino acid        sequences of SEQ ID NOs:4-6.

(47) CD19 (CD19 molecule)

Nucleotide

Genbank accession no. NM_001178098

Genbank version no. NM_001178098.1 GI:296010920

Genbank record update date: Sep. 10, 2012 12:43 AM

Polypeptide

Genbank accession no. NP_001171569

Genbank version no. NP_001171569.1 GI:296010921

Genbank record update date: Sep. 10, 2012 12:43 AM

Cross-References

Tedder T F., et al J. Immunol. 143 (2): 712-7 (1989)

Other Information

Official Symbol: CD19

Other Aliases: B4, CVID3

Other Designations: B-lymphocyte antigen CD19; B-lymphocyte surfaceantigen B4; T-cell surface antigen Leu-12; differentiation antigen CD19

Antibodies

Immunogen: HuB4—Al-Katib A M., et al Clin Cancer Res. 2009 Jun. 15;15(12):4038-45.

4G7: Kügler M., et al Protein Eng Des Sel. 2009 March; 22(3):135-47

-   -   For example, sequences in FIG. 3 of of Knappik, A. et al. J Mol        Biol 2000 February; 296(1):57-86

AstraZeneca/MedImmune: MEDI-551—Herbst R., et al J Pharmacol Exp Ther.2010 October; 335(1):213-22

Glenmark Pharmaceuticals: GBR-401—Hou S., et al Mol Cancer Ther November2011 10 (Meeting Abstract Supplement) C164

U.S. Pat. No. 7,109,304 (Immunomedics)

-   -   For example, an antibody comprising the sequence of hA19Vk (SEQ        ID NO:7) and the sequence of hA19VH (SEQ ID NO:10)

U.S. Pat. No. 7,902,338 (Immunomedics)

-   -   For example, an antibody or antigen-binding fragment thereof        that comprises the light chain complementarity determining        region CDR sequences CDR1 of SEQ ID NO: 16 (KASQSVDYDGDSYLN);        CDR2 of SEQ ID NO: 17 (DASNLVS); and CDR3 of SEQ ID NO: 18        (QQSTEDPWT) and the heavy chain CDR sequences CDR1 of SEQ ID NO:        19 (SYWMN); CDR2 of SEQ ID NO: 20 (QIWPGDGDTNYNGKFKG) and CDR3        of SEQ ID NO: 21 (RETTTVGRYYYAMDY) and also comprises human        antibody framework (FR) and constant region sequences with one        or more framework region amino acid residues substituted from        the corresponding framework region sequences of the parent        murine antibody, and wherein said substituted FR residues        comprise the substitution of serine for phenylalanine at Kabat        residue 91 of the heavy chain variable region.

Medarex: MDX-1342—Cardarelli P M., et al Cancer Immunol Immunother. 2010February; 59(2):257-65.

MorphoSys/Xencor: MOR-208/XmAb-5574—Zalevsky J., et al Blood. 2009 Apr.16; 113(16):3735-43

U.S. Pat. No. 7,968,687 (Seattle Genetics)

-   -   An antibody or antigen-binding fragment comprising a heavy chain        variable domain comprising the amino acid sequence of SEQ ID        NO:9 and a light chain variable domain comprising the amino acid        sequence of SEQ ID NO: 24.

4G7 chim—Lang P., et al Blood. 2004 May 15; 103(10):3982-5 (Universityof Tübingen)

-   -   For example, FIG. 6 and SEQ ID No: 80 of US20120082664

Zhejiang University School of Medicine: 2E8—Zhang J., et al J DrugTarget. 2010 November; 18(9):675-8

(48) IL2RA (Interleukin 2 receptor, alpha); NCBI Reference Sequence:NM_000417.2);

Nucleotide

Genbank accession no. NM_000417

Genbank version no. NM_000417.2 GI:269973860

Genbank record update date: Sep. 9, 2012 04:59 PM

Polypeptide

Genbank accession no. NP_000408

Genbank version no. NP_000408.1 GI:4557667

Genbank record update date: Sep. 9, 2012 04:59 PM

Cross-References

Kuziel W. A., et al J. Invest. Dermatol. 94 (6 SUPPL), 27S-32S (1990)

Other Information

Official Symbol: IL2RA

Other Aliases: RP11-536K7.1, CD25, IDDM10, IL2R, TCGFR

Other Designations: FIL-2 receptor subunit alpha; IL-2-RA; IL-2R subunitalpha; IL2-RA;

TAC antigen; interleukin-2 receptor subunit alpha; p55

Antibodies

U.S. Pat. No. 6,383,487 (Novartis/UCL: Baxilisimab [Simulect])

U.S. Pat. No. 6,521,230 (Novartis/UCL: Baxilisimab [Simulect])

-   -   For example, an antibody having an antigen binding site        comprises at least one domain which comprises CDR1 having the        amino acid sequence in SEQ. ID. NO: 7, CDR2 having the amino        acid sequence in SEQ. ID. NO: 8, and CDR3 chaving the amino acid        sequence in SEQ. ID. NO: 9; or said CDR1, CDR2 and CDR3 taken in        sequence as a whole comprise an amino acid sequence which is at        least 90% identical to SEQ. ID. NOs: 7, 8 and 9 taken in        sequence as a whole.

Daclizumab—Rech A J., et al Ann N Y Acad Sci. 2009 September;1174:99-106 (Roche)

(49) AXL (AXL receptor tyrosine kinase)

Nucleotide

Genbank accession no. M76125

Genbank version no. M76125.1 GI:292869

Genbank record update date: Jun. 23, 2010 08:53 AM

Polypeptide

Genbank accession no. AAA61243

Genbank version no. AAA61243.1 GI:29870

Genbank record update date: Jun. 23, 2010 08:53 AM

Cross-References

O'Bryan J. P., et al Mol. Cell. Biol. 11 (10), 5016-5031 (1991);Bergsagel P. L., et al J. Immunol. 148 (2), 590-596 (1992)

Other Information

Official Symbol: AXL

Other Aliases: JTK11, UFO

Other Designations: AXL oncogene; AXL transforming sequence/gene;oncogene AXL; tyrosine-protein kinase receptor UFO

Antibodies

YW327.6S2—Ye X., et al Oncogene. 2010 Sep. 23; 29(38):5254-64.(Genentech)

BergenBio: BGB324 (http://www.bergenbio.com/BGB324)

(50) CD30—TNFRSF8 (Tumor necrosis factor receptor superfamily, member 8)

Nucleotide

Genbank accession no. M83554

Genbank version no. M83554.1 GI:180095

Genbank record update date: Jun. 23, 2010 08:53 AM

Polypeptide

Genbank accession no. AAA51947

Genbank version no. AAA51947.1 GI:180096

Genbank record update date: Jun. 23, 2010 08:53 AM

Cross-References

Durkop H., et al Cell 68 (3), 421-427 (1992)

Other Information

Official Symbol: TNFRSF8

Other Aliases: CD30, D1S166E, Ki-1

Other Designations: CD30L receptor; Ki-1 antigen; cytokine receptorCD30; lymphocyte activation antigen CD30; tumor necrosis factor receptorsuperfamily member 8

(51) BCMA (B-cell maturation antigen)—TNFRSF17 (Tumor necrosis factorreceptor superfamily, member 17)

Nucleotide

Genbank accession no. Z29574

Genbank version no. Z29574.1 GI:471244

Genbank record update date: Feb. 2, 2011 10:40 AM

Polypeptide

Genbank accession no. CAA82690

Genbank version no. CAA82690.1 GI:471245

Genbank record update date: Feb. 2, 2011 10:40 AM

Cross-References

Laabi Y., et al Nucleic Acids Res. 22 (7), 1147-1154 (1994)

Other Information

Official Symbol: TNFRSFI7

Other Aliases: BCM, BCMA, CD269

Other Designations: B cell maturation antigen; B-cell maturation factor;B-cell maturation protein; tumor necrosis factor receptor superfamilymember 17

(52) CT Ags—CTA (Cancer Testis Antigens)

Cross-References

Fratta E., et al. Mol Oncol. 2011 April; 5(2):164-82; Lim SH., at al AmJ Blood Res. 2012; 2(1):29-35.

(53) CD174 (Lewis Y)—FUT3 (fucosyltransferase 3 (galactoside3(4)-L-fucosyltransferase, Lewis blood group)

Nucleotide

Genbank accession no. NM000149

Genbank version no. NM000149.3 GI:148277008

Genbank record update date: Jun. 26, 2012 04:49 PM

Polypeptide

Genbank accession no. NP_000140

Genbank version no. NP_000140.1 GI:4503809

Genbank record update date: Jun. 26, 2012 04:49 PM

Cross-References

Kukowska-Latallo, J. F., et al Genes Dev. 4 (8), 1288-1303 (1990)

Other Information

Official Symbol: FUT3

Other Aliases: CD174, FT3B, FucT-III, LE, Les

Other Designations: Lewis FT; alpha-(1,3/1,4)-fucosyltransferase; bloodgroup Lewis alpha-4-fucosyltransferase; fucosyltransferase III;galactoside 3(4)-L-fucosyltransferase

(54) CLEC14A (C-type lectin domain family 14, member A; Genbankaccession no. NM175060)

Nucleotide

Genbank accession no. NM175060

Genbank version no. NM175060.2 GI:371123930

Genbank record update date: Apr. 1, 2012 03:34 PM

Polypeptide

Genbank accession no. NP_778230

Genbank version no. NP_778230.1 GI:28269707

Genbank record update date: Apr. 1, 2012 03:34 PM

Other Information

Official Symbol: CLEC14A

Other Aliases: UNQ236/PRO269, C14orf27, CEG1, EGFR-5

Other Designations: C-type lectin domain family 14 member A; CIECT andEGF-like domain containing protein; epidermal growth factor receptor 5

(55) GRP78—HSPA5 (heat shock 70 kDa protein 5 (glucose-regulatedprotein, 78 kDa)

Nucleotide

Genbank accession no. NM005347

Genbank version no. NM005347.4 GI:305855105

Genbank record update date: Sep. 30, 2012 01:42 PM

Polypeptide

Genbank accession no. NP_005338

Genbank version no. NP_005338.1 GI:16507237

Genbank record update date: Sep. 30, 2012 01:42 PM

Cross-References

Ting J., et al DNA 7 (4), 275-286 (1988)

Other Information

Official Symbol: HSPA5

Other Aliases: BIP, GRP78, MIF2

Other Designations: 78 kDa glucose-regulated protein; endoplasmicreticulum lumenal Ca(2+)-binding protein grp78; immunoglobulin heavychain-binding protein

(56) CD70 (CD70 molecule) L08096

Nucleotide

Genbank accession no. L08096

Genbank version no. L08096.1 GI:307127

Genbank record update date: Jun. 23, 2012 08:54 AM

Polypeptide

Genbank accession no. AAA36175

Genbank version no. AAA36175.1 GI:307128

Genbank record update date: Jun. 23, 2012 08:54 AM

Cross-References

Goodwin R. G., et al Cell 73 (3), 447-456 (1993)

Other Information

Official Symbol: CD70

Other Aliases: CD27L, CD27LG, TNFSF7

Other Designations: CD27 ligand; CD27-L; CD70 antigen; Ki-24 antigen;surface antigen CD70; tumor necrosis factor (ligand) superfamily, member7; tumor necrosis factor ligand superfamily member 7

Antibodies

MDX-1411 against CD70 (Medarex)

h1F6 (Oflazoglu, E., et al, Clin Cancer Res. 2008 Oct. 1;14(19):6171-80; Seattle Genetics)

-   -   For example, see US20060083736 SEQ ID NOs: 1, 2, 11 and 12 and        FIG. 1.

(57) Stem Cell specific antigens. For example:

-   -   5T4 (see entry (63) below)    -   CD25 (see entry (48) above)    -   CD32        -   Polypeptide            -   Genbank accession no. ABK42161            -   Genbank version no. ABK42161.1 GI:117616286            -   Genbank record update date: Jul. 25, 2007 03:00 PM    -   LGR5/GPR49        -   Nucleotide            -   Genbank accession no. NM_003667            -   Genbank version no. NM_003667.2 GI:24475886            -   Genbank record update date: Jul. 22, 2012 03:38 PM        -   Polypeptide            -   Genbank accession no. NP_003658            -   Genbank version no. NP_003658.1 GI:4504379        -   Genbank record update date: Jul. 22, 2012 03:38 PM    -   Prominin/CD133        -   Nucleotide            -   Genbank accession no. NM_006017            -   Genbank version no. NM_006017.2 GI:224994187            -   Genbank record update date: Sep. 30, 2012 01:47 PM        -   Polypeptide            -   Genbank accession no. NP_006008            -   Genbank version no. NP_006008.1 GI:5174387            -   Genbank record update date: Sep. 30, 2012 01:47 PM

(58) ASG-5

Cross-References

(Smith L. M., et. al AACR 2010 Annual Meeting (abstract #2590); Gudas J.M., et. al. AACR 2010 Annual Meeting (abstract #4393)

Antibodies

Anti-AGS-5 Antibody: M6.131 (Smith, L. M., et. al AACR 2010 AnnualMeeting (abstract #2590)

(59) ENPP3 (Ectonucleotide pyrophosphatase/phosphodiesterase 3)

Nucleotide

Genbank accession no. AF005632

Genbank version no. AF005632.2 GI:4432589

Genbank record update date: Mar. 10, 2010 09:41 PM

Polypeptide

Genbank accession no. AAC51813

Genbank version no. AAC51813.1 GI:2465540

Genbank record update date: Mar. 10, 2010 09:41 PM

Cross-References

Jin-Hua P., et al Genomics 45 (2), 412-415 (1997)

Other Information

Official Symbol: ENPP3

Other Aliases: RP5-988G15.3, B10, CD203c, NPP3, PD-IBETA, PDNP3

Other Designations: E-NPP 3; dJ1005H11.3 (phosphodiesterase I/nucleotidepyrophosphatase 3); dJ914N13.3 (phosphodiesterase I/nucleotidepyrophosphatase 3); ectonucleotide pyrophosphatase/phosphodiesterasefamily member 3; gp13ORB13-6; phosphodiesterase I beta;phosphodiesterase I/nucleotide pyrophosphatase 3; phosphodiesterase-Ibeta

(60) PRR4 (Proline rich 4 (lacrimal))

Nucleotide

Genbank accession no. NM_007244

Genbank version no. NM_007244.2 GI:154448885

Genbank record update date: Jun. 28, 2012 12:39 PM

Polypeptide

Genbank accession no. NP_009175

Genbank version no. NP_009175.2 GI:154448886

Genbank record update date: Jun. 28, 2012 12:39 PM

Cross-References

Dickinson D. P., et al Invest. Ophthalmol. Vis. Sci. 36 (10), 2020-2031(1995)

Other Information

Official Symbol: PRR4

Other Aliases: LPRP, PROL4

Other Designations: lacrimal proline-rich protein; nasopharyngealcarcinoma-associated proline-rich protein 4; proline-rich polypeptide 4;proline-rich protein 4

(61) GCC—GUCY2C (guanylate cyclase 2C (heat stable enterotoxin receptor)

Nucleotide

Genbank accession no. NM_004963

Genbank version no. NM_004963.3 GI:222080082

Genbank record update date: Sep. 2, 2012 01:50 PM

Polypeptide

Genbank accession no. NP_004954

Genbank version no. NP_004954.2 GI:222080083

Genbank record update date: Sep. 2, 2012 01:50 PM

Cross-References

De Sauvage F. J., et al J. Biol. Chem. 266 (27), 17912-17918 (1991);Singh S., et al Biochem. Biophys. Res. Commun. 179 (3), 1455-1463 (1991)

Other Information

Official Symbol: GUCY2C

Other Aliases: DIAR6, GUC2C, MUCIL, STAR

Other Designations: GC-C; STA receptor; guanylyl cyclase C; hSTAR;heat-stable enterotoxin receptor; intestinal guanylate cyclase

(62) Liv-1—SLC39A6 (Solute carrier family 39 (zinc transporter), member6)

Nucleotide

Genbank accession no. U41060

Genbank version no. U41060.2 GI:12711792

Genbank record update date: Nov. 30, 2009 04:35 PM

Polypeptide

Genbank accession no. AAA96258

Genbank version no. AAA96258.2 GI:12711793

Genbank record update date: Nov. 30, 2009 04:35 PM

Cross-References

Taylor K M., et al Biochim Biophys Acta. 2003 Apr. 1; 1611(1-2):16-30

Other Information

Official Symbol: SLC39A6

Other Aliases: LIV-1

Other Designations: LIV-1 protein, estrogen regulated; ZIP-6;estrogen-regulated protein LIV-1; solute carrier family 39 (metal iontransporter), member 6; solute carrier family 39 member 6; zinctransporter ZIP6; zrt- and lrt-like protein 6

(63) 5T4, Trophoblast glycoprotein, TPBG—TPBG (trophoblast glycoprotein)

Nucleotide

Genbank accession no. AJ012159

Genbank version no. AJ012159.1 GI:3805946

Genbank record update date: Feb. 1, 2011 10:27 AM

Polypeptide

Genbank accession no. CAA09930

Genbank version no. CAA09930.1 GI:3805947

Genbank record update date: Feb. 1, 2011 10:27 AM

Cross-References

King K. W., et al Biochim. Biophys. Acta 1445 (3), 257-270 (1999)

Other Information

-   -   Official Symbol: TPBG    -   Other Aliases: 5T4, 5T4AG, M6P1    -   Other Designations: 5T4 oncofetal antigen; 5T4 oncofetal        trophoblast glycoprotein; 5T4 oncotrophoblast glycoprotein

(64) CD56—NCMA1 (Neural cell adhesion molecule 1)

Nucleotide

Genbank accession no. NM_000615

Genbank version no. NM_000615.6 GI:336285433

Genbank record update date: Sep. 23, 2012 02:32 PM

Polypeptide

Genbank accession no. NP_000606

Genbank version no. NP_000606.3 GI:94420689

Genbank record update date: Sep. 23, 2012 02:32 PM

Cross-References

Dickson, G., et al, Cell 50 (7), 1119-1130 (1987)

Other Information

Official Symbol: NCAM1

Other Aliases: CD56, MSK39, NCAM

Other Designations: antigen recognized by monoclonal antibody 5.1H11;neural cell adhesion molecule, NCAM

Antibodies

Immunogen: HuN901 (Smith S V., et al Curr Opin Mol Ther. 2005 August;7(4):394-401)

-   -   For example, see humanized from murine N901 antibody. See FIGS.        1b and 1e of Roguska, M. A., et al. Proc Natl Acad Sci USA        February 1994; 91:969-973.

(65) CanAg (Tumor associated antigen CA242)

Cross-References

Haglund C., et al Br J Cancer 60:845-851, 1989; Baeckstrom D., et al JBiol Chem 266:21537-21547, 1991

Antibodies

huC242 (Tolcher A W et al., J Clin Oncol. 2003 January 15; 21(2):211-22;Immunogen)

-   -   For example, see US20080138898A1 SEQ ID NO: 1 and 2

(66) FOLR1 (Folate Receptor 1)

Nucleotide

Genbank accession no. J05013

Genbank version no. J05013.1 GI:182417

Genbank record update date: Jun. 23, 2010 08:47 AM

Polypeptide

Genbank accession no. AAA35823

Genbank version no. AAA35823.1 GI:182418

Genbank record update date: Jun. 23, 2010 08:47 AM

Cross-References

Elwood P.C., et al J. Biol. Chem. 264 (25), 14893-14901 (1989)

Other Information

Official Symbol: FOLR1

Other Aliases: FBP, FOLR

Other Designations: FR-alpha; KB cells FBP; adult folate-bindingprotein; folate binding protein; folate receptor alpha; folate receptor,adult; ovarian tumor-associated antigen MOv18

Antibodies

M9346A—Whiteman K R., et al Cancer Res Apr. 15, 2012; 72(8 Supplement):4628 (Immunogen)

(67) GPNMB (Glycoprotein (transmembrane) nmb)

Nucleotide

Genbank accession no. X76534

Genbank version no. X76534.1 GI:666042

Genbank record update date: Feb. 2, 2011 10:10 AM

Polypeptide

Genbank accession no. CAA54044

Genbank version no. CAA54044.1 GI:666043

Genbank record update date: Feb. 2, 2011 10:10 AM

Cross-References

Weterman M. A., et al Int. J. Cancer 60 (1), 73-81 (1995)

Other Information

Official Symbol: GPNMB

Other Aliases: UNQ1725/PRO9925, HGFIN, NMB

Other Designations: glycoprotein NMB; glycoprotein nmb-like protein;osteoactivin; transmembrane glycoprotein HGFIN; transmembraneglycoprotein NMB

Antibodies

Celldex Therapeutics: CR011 (Tse K F., et al Clin Cancer Res. 2006 Feb.15; 12(4):1373-82)

-   -   For example, see EP1827492B1 SEQ ID NO: 22, 24, 26, 31, 33 and        35

(68) TIM-1—HAVCR1 (Hepatitis A virus cellular receptor 1)

Nucleotide

Genbank accession no. AF043724

Genbank version no. AF043724.1 GI:2827453

Genbank record update date: Mar. 10, 2010 06:24 PM

Polypeptide

Genbank accession no. AAC39862

Genbank version no. AAC39862.1 GI:2827454

Genbank record update date: Mar. 10, 2010 06:24 PM

Cross-References

Feigelstock D., et al J. Virol. 72 (8), 6621-6628 (1998)

Other Information

Official Symbol: HAVCR1

Other Aliases: HAVCR, HAVCR-1, KIM-1, KIM1, TIM, TIM-1, TIM1, TIMD-1,TIMD1

Other Designations: T cell immunoglobin domain and mucin domain protein1; T-cell membrane protein 1; kidney injury molecule 1

(69) RG-1/Prostate tumor target Mindin—Mindin/RG-I

Cross-References

Parry R., et al Cancer Res. 2005 Sep. 15; 65(18):8397-405

(70) 87-H4—VTCN1 (V-set domain containing T cell activation inhibitor 1

Nucleotide

Genbank accession no. BX648021

Genbank version no. BX648021.1 GI:34367180

Genbank record update date: Feb. 2, 2011 08:40 AM

Cross-References

Sica G L., et al Immunity. 2003 June; 18(6):849-61

Other Information

Official Symbol: VTCN1

Other Aliases: RP11-229A19.4, B7-H4, B7H4, B7S1, B7X, B7h.5, PRO1291,VCTN1

Other Designations: B7 family member, H4; B7 superfamily member 1; Tcell costimulatory molecule B7x; T-cell costimulatory molecule B7x;V-set domain-containing T-cell activation inhibitor 1; immunecostimulatory protein B7-H4

(71) PTK7 (PTK7 protein tyrosine kinase 7)

Nucleotide

Genbank accession no. AF447176

Genbank version no. AF447176.1 GI:17432420

Genbank record update date: Nov. 28, 2008 01:51 PM

Polypeptide

Genbank accession no. AAL39062

Genbank version no. AAL39062.1 GI:17432421

Genbank record update date: Nov. 28, 2008 01:51 PM

Cross-References

Park S. K., et al J. Biochem. 119 (2), 235-239 (1996)

Other Information

Official Symbol: PTK7

Other Aliases: CCK-4, CCK4

Other Designations: colon carcinoma kinase 4; inactive tyrosine-proteinkinase 7; pseudo tyrosine kinase receptor 7; tyrosine-proteinkinase-like 7 (72) CD37 (CD37 molecule)

Nucleotide

Genbank accession no. NM_001040031

Genbank version no. NM_001040031.1 GI:91807109

Genbank record update date: Jul. 29, 2012 02:08 PM

Polypeptide

Genbank accession no. NP_001035120

Genbank version no. NP_001035120.1 GI:91807110

Genbank record update date: Jul. 29, 2012 02:08 PM

Cross-References

Schwartz-Albiez R., et al J. Immunol. 140 (3), 905-914 (1988)

Other Information

Official Symbol: CD37

Other Aliases: GP52-40, TSPAN26

Other Designations: CD37 antigen; cell differentiation antigen 37;leukocyte antigen CD37; leukocyte surface antigen CD37; tetraspanin-26;tspan-26

Antibodies

Boehringer Ingelheim: mAb 37.1 (Heider K H., et al Blood. 2011 Oct. 13;118(15):4159-68)

Trubion: CD37-SMIP (G28-1 scFv-Ig) ((Zhao X., et al Blood. 2007; 110:2569-2577)

-   -   For example, see US20110171208A1 SEQ ID NO: 253

Immunogen: K7153A (Deckert J., et al Cancer Res Apr. 15, 2012; 72(8Supplement): 4625)

(73) CD138—SDC1 (syndecan 1)

Nucleotide

Genbank accession no. AJ551176

Genbank version no. AJ551176.1 GI:29243141

Genbank record update date: Feb. 1, 2011 12:09 PM

Polypeptide

Genbank accession no. CAD80245

Genbank version no. CAD80245.1 GI:29243142

Genbank record update date: Feb. 1, 2011 12:09 PM

Cross-References

O'Connell F P., et al Am J Clin Pathol. 2004 February; 121(2):254-63

Other Information

Official Symbol: SDC1

Other Aliases: CD138, SDC, SYND1, syndecan

Other Designations: CD138 antigen; heparan sulfate proteoglycanfibroblast growth factor receptor; syndecan proteoglycan 1; syndecan-1

Antibodies

Biotest: chimerized MAb (nBT062)—(Jagannath S., et al Poster ASH #3060,2010; WIPO Patent Application WO/2010/128087)

-   -   For example, see US20090232810 SEQ ID NO: 1 and 2

Immunogen: B-B4 (Tassone P., et al Blood 104_3688-3696)

-   -   For example, see US20090175863A1 SEQ ID NO: 1 and 2

(74) CD74 (CD74 molecule, major histocompatibility complex, class IIinvariant chain)

Nucleotide

Genbank accession no. NM_004355

Genbank version no. NM_004355.1 GI:343403784

Genbank record update date: Sep. 23, 2012 02:30 PM

Polypeptide

Genbank accession no. NP_004346

Genbank version no. NP_004346.1 GI:10835071

Genbank record update date: Sep. 23, 2012 02:30 PM

Cross-References

Kudo, J., et al Nucleic Acids Res. 13 (24), 8827-8841 (1985)

Other Information

Official Symbol: CD74

Other Aliases: DHLAG, HLADG, II, Ia-GAMMA

Other Designations: CD74 antigen (invariant polypeptide of majorhistocompatibility complex, class II antigen-associated); HLA class IIhistocompatibility antigen gamma chain; HLA-DR antigens-associatedinvariant chain; HLA-DR-gamma; la-associated invariant chain; MHC HLA-DRgamma chain; gamma chain of class II antigens; p33

Antibodies

Immunomedics: hLL1 (Milatuzumab,)—Berkova Z., et al Expert Opin InvestigDrugs. 2010 January; 19(1):141-9)

-   -   For example, see US20040115193 SEQ ID NOs: 19, 20, 21, 22, 23        and 24

Genmab: HuMax-CD74 (see website)

(75) Claudins—CLs (Claudins)

Cross-References

Offner S., et al Cancer Immunol Immunother. 2005 May; 54(5):431-45,Suzuki H., et al Ann N Y Acad Sci. 2012 July; 1258:65-70)

In humans, 24 members of the family have been described—see literaturereference.

(76) EGFR (Epidermal growth factor receptor)

Nucleotide

Genbank accession no. NM_005228

Genbank version no. NM_005228.3 GI:41927737

Genbank record update date: Sep. 30, 2012 01:47 PM

Polypeptide

Genbank accession no. NP_005219

Genbank version no. NP_005219.2 GI:29725609

Genbank record update date: Sep. 30, 2012 01:47 PM

Cross-References

Dhomen N S., et al Crit Rev Oncog. 2012; 17(1):31-50

Other Information

Official Symbol: EGFR

Other Aliases: ERBB, ERBB1, HER1, PIG61, mENA

Other Designations: avian erythroblastic leukemia viral (v-erb-b)oncogene homolog; cell growth inhibiting protein 40; cellproliferation-inducing protein 61; proto-oncogene c-ErbB-1; receptortyrosine-protein kinase erbB-1

Antibodies

BMS: Cetuximab (Erbitux)—Broadbridge V T., et al Expert Rev AnticancerTher. 2012 May; 12(5):555-65.

-   -   For example, see U.S. Pat. No. 6,217,866—ATTC deposit No. 9764.

Amgen: Panitumumab (Vectibix)—Argiles G., et al Future Oncol. 2012April; 8(4):373-89

-   -   For example, see U.S. Pat. No. 6,235,883 SEQ ID NOs: 23-38.

Genmab: Zalutumumab—Rivera F., et al Expert Opin Biol Ther. 2009 May;9(5):667-74.

Y M Biosciences: Nimotuzumab—Ramakrishnan M S., et al MAbs. 2009January-February; 1(1):41-8.

-   -   For example, see U.S. Pat. No. 5,891,996 SEQ ID NOs: 27-34.

(77) Her3 (ErbB3)—ERBB3 (v-erb-b2 erythroblastic leukemia viral oncogenehomolog 3 (avian))

Nucleotide

Genbank accession no. M34309

Genbank version no. M34309.1 GI:183990

Genbank record update date: Jun. 23, 2010 08:47 PM

Polypeptide

Genbank accession no. AAA35979

Genbank version no. AAA35979.1 GI:306841

Genbank record update date: Jun. 23, 2010 08:47 PM

Cross-References

Plowman, G. D., et al., Proc. Natl. Acad. Sci. U.S.A. 87 (13), 4905-4909(1990)

Other Information

Official Symbol: ERBB3

Other Aliases: ErbB-3, HER3, LCCS2, MDA-BF-1, c-erbB-3, c-erbB3,erbB3-S, p180-ErbB3, p45-sErbB3, p85-sErbB3

Other Designations: proto-oncogene-like protein c-ErbB-3; receptortyrosine-protein kinase erbB-3; tyrosine kinase-type cell surfacereceptor HER3

Antibodies

Merimack Pharma: MM-121 (Schoeberl B., et al Cancer Res. 2010 Mar. 15;70(6):2485-2494)

-   -   For example, see US2011028129 SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7        and 8.

(78) RON—MST1R (macrophage stimulating 1 receptor (c-met-relatedtyrosine kinase))

Nucleotide

Genbank accession no. X70040

Genbank version no. X70040.1 GI:36109

Genbank record update date: Feb. 2, 2011 10:17 PM

Polypeptide

Genbank accession no. CCA49634

Genbank version no. CCA49634.1 GI:36110

Genbank record update date: Feb. 2, 2011 10:17 PM

Cross-References

Ronsin C., et al Oncogene 8 (5), 1195-1202 (1993)

Other Information

Official Symbol: MST1R

Other Aliases: CD136, CDw136, PTK8, RON

Other Designations: MSP receptor; MST1R variant RON30; MST1R variantRON62; PTK8 protein tyrosine kinase 8; RON variant E2E3; c-met-relatedtyrosine kinase; macrophage-stimulating protein receptor; p185-Ron;soluble RON variant 1; soluble RON variant 2; soluble RON variant 3;soluble RONvariant 4

(79) EPHA2 (EPH receptor A2)

Nucleotide

Genbank accession no. BC037166

Genbank version no. BC037166.2 GI:33879863

Genbank record update date: Mar. 6, 2012 01:59 PM

Polypeptide

Genbank accession no. AAH37166

Genbank version no. AAH37166.1 GI:22713539

Genbank record update date: Mar. 6, 2012 01:59 PM

Cross-References

Strausberg R. L., et al Proc. Natl. Acad. Sci. U.S.A. 99 (26),16899-16903 (2002)

Other Information

Official Symbol: EPHA2

Other Aliases: ARCC2, CTPA, CTPP1, ECK

Other Designations: ephrin type-A receptor 2; epithelial cell receptorprotein tyrosine kinase; soluble EPHA2 variant 1; tyrosine-proteinkinase receptor ECK

Antibodies

Medimmune: 1C1 (Lee J W., et al Clin Cancer Res. 2010 May 1;16(9):2562-2570)

-   -   For example, see US20090304721A1 FIGS. 7 and 8.

(80) CD20—MS4A1 (membrane-spanning 4-domains, subfamily A, member 1)

Nucleotide

Genbank accession no. M27394

Genbank version no. M27394.1 GI:179307

Genbank record update date: Nov. 30, 2009 11:16 AM

Polypeptide

Genbank accession no. AAA35581

Genbank version no. AAA35581.1 GI:179308

Genbank record update date: Nov. 30, 2009 11:16 AM

Cross-References

Tedder T. F., et al Proc. Natl. Acad. Sci. U.S.A. 85 (1), 208-212 (1988)

Other Information

Official Symbol: MS4A1

Other Aliases: B1, Bp35, CD20, CVID5, LEU-16, MS4A2, S7

Other Designations: B-lymphocyte antigen CD20; B-lymphocyte cell-surfaceantigen B1; CD20 antigen; CD20 receptor; leukocyte surface antigenLeu-16

Antibodies

Genentech/Roche: Rituximab—Abdulla N E., et al BioDrugs. 2012 Apr. 1;26(2):71-82.

For example, see U.S. Pat. No. 5,736,137, ATCC deposit No. HB-69119.

GSK/Genmab: Ofatumumab—Nightingale G., et al Ann Pharmacother. 2011October; 45(10):1248-55.

-   -   For example, see US20090169550A1 SEQ ID NOs: 2, 4 and 5.

Immunomedics: Veltuzumab-Goldenberg D M., et al Leuk Lymphoma. 2010 May;51(5):747-55.

-   -   For example, see U.S. Pat. No. 7,919,273B2 SEQ ID NOs: 1, 2, 3,        4, 5 and 6.

(81) Tenascin C—TNC (Tenascin C)

Nucleotide

Genbank accession no. NM_002160

Genbank version no. NM_002160.3 GI:340745336

Genbank record update date: Sep. 23, 2012 02:33 PM

Polypeptide

Genbank accession no. NP_002151

Genbank version no. NP_002151.2 GI:153946395

Genbank record update date: Sep. 23, 2012 02:33 PM

Cross-References

Nies D. E., et al J. Biol. Chem. 266 (5), 2818-2823 (1991); Siri A., etal Nucleic Acids Res. 19 (3), 525-531 (1991)

Other Information

Official Symbol: TNC

Other Aliases: 150-225, GMEM, GP, HXB, JI, TN, TN-C

Other Designations: GP 150-225; cytotactin;glioma-associated-extracellular matrix antigen; hexabrachion (tenascin);myotendinous antigen; neuronectin; tenascin; tenascin-C isoform14/AD1/16

Antibodies

Philogen: G11 (von Lukowicz T., et al J Nucl Med. 2007 April;48(4):582-7) and F16 (Pedretti M., et al Lung Cancer. 2009 April;64(1):28-33)

-   -   For example, see U.S. Pat. No. 7,968,685 SEQ ID NOs: 29, 35, 45        and 47.

(82) FAP (Fibroblast activation protein, alpha)

Nucleotide

Genbank accession no. U09278

Genbank version no. U09278.1 GI:1888315

Genbank record update date: Jun. 23, 2010 09:22 AM

Polypeptide

Genbank accession no. AAB49652

Genbank version no. AAB49652.1 GI:1888316

Genbank record update date: Jun. 23, 2010 09:22 AM

Cross-References

Scanlan, M. J., et al Proc. Natl. Acad. Sci. U.S.A. 91 (12), 5657-5661(1994)

Other Information

Official Symbol: FAP

Other Aliases: DPPIV, FAPA

Other Designations: 170 kDa melanoma membrane-bound gelatinase; integralmembrane serine protease; seprase

(83) DKK-1 (Dickkopf 1 homolog (Xenopus laevis)

Nucleotide

Genbank accession no. NM_012242

Genbank version no. NM_012242.2 GI:61676924

Genbank record update date: Sep. 30, 2012 01:48 PM

Polypeptide

Genbank accession no. NP_036374

Genbank version no. NP_036374.1 GI:7110719

Genbank record update date: Sep. 30, 2012 01:48 PM

Cross-References

Fedi P. et al J. Biol. Chem. 274 (27), 19465-19472 (1999)

Other Information

Official Symbol: DKK1

Other Aliases: UNQ492/PRO1008, DKK-1, SK

Other Designations: dickkopf related protein-1; dickkopf-1 like;dickkopf-like protein 1; dickkopf-related protein 1; hDkk-1

Antibodies

Novartis: BHQ880 (Fulciniti M., et al Blood. 2009 Jul. 9;114(2):371-379)

-   -   For example, see US20120052070A1 SEQ ID NOs: 100 and 108.

(84) CD52 (CD52 molecule)

Nucleotide

Genbank accession no. NM_001803

Genbank version no. NM_001803.2 GI:68342029

Genbank record update date: Sep. 30, 2012 01:48 PM

Polypeptide

Genbank accession no. NP_001794

Genbank version no. NP_001794.2 GI:68342030

Genbank record update date: Sep. 30, 2012 01:48 PM

Cross-References

Xia M. Q., et al Eur. J. Immunol. 21 (7), 1677-1684 (1991)

Other Information

Official Symbol: CD52

Other Aliases: CDW52

Other Designations: CAMPATH-1 antigen; CD52 antigen (CAMPATH-1 antigen);CDW52 antigen (CAMPATH-1 antigen); cambridge pathology 1 antigen;epididymal secretory protein E5; he5; human epididymis-specific protein5

Antibodies

Alemtuzumab (Campath)—Skoetz N., et al Cochrane Database Syst Rev. 2012Feb. 15; 2:CD008078.

For example, see Drugbank Acc. No. DB00087 (BIOD00109, BTD00109) (85)CS1-SLAMF7 (SLAM family member 7)

Nucleotide

Genbank accession no. NM_021181

Genbank version no. NM_021181.3 GI:1993571

Genbank record update date: Jun. 29, 2012 11:24 AM

Polypeptide

Genbank accession no. NP_067004

Genbank version no. NP_067004.3 GI:19923572

Genbank record update date: Jun. 29, 2012 11:24 AM

Cross-References

Boles K. S., et al Immunogenetics 52 (3-4), 302-307 (2001)

Other Information

Official Symbol: SLAMF7

Other Aliases: UNQ576/PRO1138, 19A, CD319, CRACC, CS1

Other Designations: 19A24 protein; CD2 subset 1; CD2-like receptoractivating cytotoxic cells; CD2-like receptor-activating cytotoxiccells; membrane protein FOAP-12; novel LY9 (lymphocyte antigen 9) likeprotein; protein 19A

Antibodies

BMS: elotuzumab/HuLuc63 (Benson D M., et al J Clin Oncol. 2012 June 1;30(16):2013-2015)

-   -   For example, see US20110206701 SEQ ID NOs: 9, 10, 11, 12, 13,        14, 15 and 16.

(86) Endoglin—ENG (Endoglin)

Nucleotide

Genbank accession no. AF035753

Genbank version no. AF035753.1 GI:3452260

Genbank record update date: Mar. 10, 2010 06:36 PM

Polypeptide

Genbank accession no. AAC32802

Genbank version no. AAC32802.1 GI:3452261

Genbank record update date: Mar. 10, 2010 06:36 PM

Cross-References

Rius C., et al Blood 92 (12), 4677-4690 (1998)

Official Symbol: ENG

Other Information

Other Aliases: RP11-228B15.2, CD105, END, HHT1, ORW, ORW1

Other Designations: CD105 antigen

(87) Annexin A1—ANXA1 (Annexin A1)

Nucleotide

Genbank accession no. X05908

Genbank version no. X05908.1 GI:34387

Genbank record update date: Feb. 2, 2011 10:02 AM

Polypeptide

Genbank accession no. CCA29338

Genbank version no. CCA29338.1 GI:34388

Genbank record update date: Feb. 2, 2011 10:02 AM

Cross-References

Wallner B. P., et al Nature 320 (6057), 77-81 (1986)

Other Information

Official Symbol: ANXA1

Other Aliases: RP11-71A24.1, ANX1, LPC1

Other Designations: annexin I (lipocortin I); annexin-1; calpactin II;calpactin-2; chromobindin-9; lipocortin I; p35; phospholipase A2inhibitory protein

(88) V-CAM (CD106)—VCAM1 (Vascular cell adhesion molecule 1)

Nucleotide

Genbank accession no. M60335

Genbank version no. M60335.1 GI:340193

Genbank record update date: Jun. 23, 2010 08:56 AM

Polypeptide

Genbank accession no. AAA61269

Genbank version no. AAA61269.1 GI:340194

Genbank record update date: Jun. 23, 2010 08:56 AM

Cross-References

Hession C., et al J. Biol. Chem. 266 (11), 6682-6685 (1991)

Other Information

Official Symbol VCAM1

Other Aliases: CD106, INCAM-100

Other Designations: CD106 antigen; vascular cell adhesion protein 1

Antibody Sequences Anti-Integrin αvβ6 RHAB6.2 (SEQ ID NO: 5)QVQLVQSGSELKKPGASVKISCKASGFAFTDSYMHWVRQAPGQGLEWMGWIDPENGDTEYAPKFQGRFVFSLDTSVSTAYLQISSLKAEDTAVYYCTRGTPTAVPNLRGDLQVLAQKVAGPYPFDYWGQGTLVTVSS RHCB6.2 (SEQ ID NO: 6)QVQLVQSGAEVKKPGASVKVSCKASGYTFIDSYMHWVRQAPGQRLEWMGWIDPENGDTEYAPKFQGRVTITTDTSASTAYMELSSLRSEDTAVYYCARGTPTAVPNLRGDLQVLAQKVAGPYPFDYWGQGTLVTVSS RHF (SEQ ID NO: 7)QVQLVQSGAEVKKPGASVKVSCKASGFNFIDSYMHWVRQAPGQRLEWMGWIDPENGDTEYAPKFQGRVTFTTDTSASTAYMELSSLRSEDTAVYYCNEGTPTGPYYFDYWGQGTLV TVSS RHFB6(SEQ ID NO: 8) QVQLVQSGAEVKKPGASVKVSCKASGFNFIDSYMHWVRQAPGQRLEWMGWIDPENGDTEYAPKFQGRVTFTTDTSASTAYMELSSLRSEDTAVYYCNEGTPTAVPNLRGDLQVLAQKVAGPYYFDYWGQGTLVTVSS RHAY100bP (SEQ ID NO: 9)QVQLVQSGSELKKPGASVKISCKASGFAFTDSYMHWVRQAPGQGLEWMGWIDPENGDTEYAPKFQGRFVFSLDTSVSTAYLQISSLKAEDTAVYYCTRGTPTGPYPFDYWGQGTLVTV SS RKF(SEQ ID NO: 10)ENVLTQSPGTLSLSPGERATLSCSASSSVSYMHWFQQKPGQAPRLLIYSTSNLASGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQRSSYPLTFGGGTKVEIK RKFL36L50 (SEQ ID NO: 11)ENVLTQSPGTLSLSPGERATLSCSASSSVSYMHWLQQKPGQAPRLLIYLTSNLASGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQRSSYPLTFGGGTKVEIK RKC (SEQ ID NO: 12)EIVLTQSPGTLSLSPGERATLSCSASSSVSYMHWFQQKPGQAPRLLIYSTSNLASGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQRSSYPLTFGGGTKVEIK Anti-CD33 CD33 Hum195 VH(SEQ ID NO: 13)QVQLVQSGAEVKKPGSSVKVSCKASGYTFTDYNMHWVRQAPGQGLEWIGYIYPYNGGTGYNQKFKSKATITADESTNTAYMELSSLRSEDTAVYYCARGRPAMDYWGQGTLVTVSSCD33 Hum195 VK (SEQ ID NO: 14)DIQMTQSPSSLSASVGDRVTITCRASESVDNYGISFMNWFQQKPGKAPKLLIYAASNQGSGVPSRFSGSGSGTDFTLTISSLQPDDFATYYCQQSKEVPWTFGQGTKVEIK Anti-CD19CD19 B4 resurfaced VH (SEQ ID NO: 15)QVQLVQPGAEVVKPGASVKLSCKTSGYTFTSNWMHWVKQRPGQGLEWIGEIDPSDSYTNYNQNFKGKAKLTVDKSTSTAYMEVSSLRSDDTAVYYCARGSNPYYYAMDYWGQGTSV TVSSCD19 B4 resurfaced VK (SEQ ID NO: 16)EIVLTQSPAIMSASPGERVTMTCSASSGVNYMHWYQQKPGTSPRRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMEPEDAATYYCHQRGSYTFGGGTKLEIK Anti-Her2Herceptin VH chain (SEQ ID NO: 1)EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVT VSSHerceptin VL chain (SEQ ID NO: 2)DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIK Anti-CD25Simulect VK (also known as Basiliximab) (SEQ ID NO: 17)QIVSTQSPAIMSASPGEKVTMTCSASSSRSYMQWYQQKPGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCHQRSSYTFGGGTKLEIK Simulect VH(SEQ ID NO: 18)QLQQSGTVLARPGASVKMSCKASGYSFTRYWMHWIKQRPGQGLEWIGAIYPGNSDTSYNQKFEGKAKLTAVTSASTAYMELSSLTHEDSAVYYCSRDYGYYFDFWGQGTTLTVSS Anti-PSMADeimmunised VH ′1 (SEQ ID NO: 19)EVQLVQSGPEVKKPGATVKISCKTSGYTFTEYTIHWVKQAPGKGLEWIGNINPNNGGTTYNQKFEDKATLTVDKSTDTAYMELSSLRSEDTAVYYCAAGWNFDYWGQGTLLTVSSDeimmunised VK ′1 (SEQ ID NO: 20)DIQMTQSPSSLSTSVGDRVTLTCKASQDVGTAVDWYQQKPGPSPKLLIYWASTRHTGIPSRFSGSGSGTDFTLTISSLQPEDFADYYCQQYNSYPLTFGPGTKVDIK Deimmunised VH1 ′5(SEQ ID NO: 21)EVKLVESGGGLVQPGGSMKLSCVASGFTFSNYWMNWVRQAPGKGLEWVAEIRSQSNNFATHYAESVKGRVTISRDDSKSIVYLQMNNLRAEDTGVYYCTRRWNNFWGQGTTVTVSSDeimmunised VH2 ′5 (SEQ ID NO: 22)EVKLVESGGGLVQPGGSLKLSCVASGFTFSNYWMNWVRQAPGKGLEWVAEIRSQSNNFATHYAESVKGRVTISRDDSKSIVYLQMNNLRAEDTAVYYCTRRWNNFWGQGTTVTVSSDeimmunised VH3 ′5 (SEQ ID NO: 23)EVQLVESGGGLVQPGGSLKLSCVASGFTFSNYWMNWVRQAPGKGLEWVAEIRSQSNNFATHYAESVKGRVTISRDDSKSIVYLQMNNLRAEDTAVYYCTRRWNNFWGQGTTVTVSSDeimmunised VH4 ′5 (SEQ ID NO: 24)EVQLVESGGGLVQPGGSLKLSCVASGFTFSNYWMNWVRQAPGKGLEWVAEIRSQSNNFATHYAESVKGRFTISRDDSKSIVYLQMNNLRAEDTAVYYCTRRWNNFWGQGTTVTVSSDeimmunised VK1 ′5 (SEQ ID NO: 25)NIVMTQFPSSMSASVGDRVTITCKASENVGTYVSWYQQKPDQSPKMLIYGASNRFTGVPDRFTGSGSATDFTLTISSLQTEDLADYYCGQSYTFPYTFGQGTKLEMK Deimmunised VK2 ′5(SEQ ID NO: 26)NIVMTQFPSSMSASVGDRVTITCKASENVGTYVSWYQQKPDQSPKMLIYGASNRFTGVPDRFSGSGSGTDFTLTISSLQAEDLADYYCGQSYTFPYTFGQGTKLEIK Deimmunised VK3 ′5(SEQ ID NO: 27)NIQMTQFPSAMSASVGDRVTITCKASENVGTYVSWYQQKPDQSPKMLIYGASNRFTGVPDRFSGSGSGTDFTLTISSLQAEDLADYYCGQSYTFPYTFGQGTKLEIK Deimmunised VK4 ′5(SEQ ID NO: 28)NIQMTQFPSAMSASVGDRVTITCKASENVGTYVSWYQQKPDQSPKMLIYGASNRFTGVPDRFSGSGSGTDFTLTISSLQAEDEADYYCGQSYTFPYTFGQGTKLEIK Deimmunised VK DI ′5(SEQ ID NO: 29)NIVMTQFPKSMSASAGERMTLTCKASENVGTYVSWYQQKPTQSPKMLIYGASNRFTGVPDRFSGSGSGTDFILTISSVQAEDLVDYYCGQSYTFPYTFGGGTKLEMK Deimmunised VH DI ′5(SEQ ID NO: 30)EVKLEESGGGLVQPGGSMKISCVASGFTFSNYWMNWVRQSPEKGLEWVAEIRSQSNNFATHYAESVKGRVIISRDDSKSSVYLQMNSLRAEDTAVYYCTRRWNNFWGQGTTVTVSSHumanised RHA ′5 (SEQ ID NO: 31)EVQLVESGGGLVQPGGSLKLSCAASGFTFSNYWMNWVRQASGKGLEWVGEIRSQSNNFATHYAESVKGRFTISRDDSKNTAYLQMNSLKTEDTAVYYCTRRWNNFWGQGTTVTVSSHumanised RHB ′5 (SEQ ID NO: 32)EVKLVESGGGLVQPGGSLKLSCAASGFTFSNYWMNWVRQASGKGLEWVAEIRSQSNNFATHYAESVKGRVIISRDDSKNTVYLQMNSLRTEDTAVYYCTRRWNNFWGQGTTVTVSSHumanised RHC ′5 (SEQ ID NO: 33)EVQLVESGGGLVQPGGSLKLSCAASGFTFSNYWMNWVRQASGKGLEWVAEIRSQSNNFATHYAESVKGRVIISRDDSKNTVYLQMNSLRTEDTAVYYCTRRWNNFWGQGTTVTVSSHumanised RHD ′5 (SEQ ID NO: 34)EVKLVESGGGLVQPGGSLKLSCAASGFTFSNYWMNWVRQASGKGLEWVGEIRSQSNNFATHYAESVKGRVIISRDDSKNTVYLQMNSLRTEDTAVYYCTRRWNNFWGQGTTVTVSSHumanised RHE ′5 (SEQ ID NO: 35)EVKLVESGGGLVQPGGSLKLSCAASGFTFSNYWMNWVRQASGKGLEWVAEIRSQSNNFATHYAESVKGRFTISRDDSKNTVYLQMNSLRTEDTAVYYCTRRWNNFWGQGTTVTVSSHumanised RHF ′5 (SEQ ID NO: 36)EVKLVESGGGLVQPGGSLKLSCAASGFTFSNYWMNWVRQASGKGLEWVAEIRSQSNNFATHYAESVKGRVIISRDDSKNTAYLQMNSLRTEDTAVYYCTRRWNNFWGQGTTVTVSSHumanised RHG ′5 (SEQ ID NO: 37)EVKLVESGGGLVQPGGSLKLSCAASGFTFSNYWMNWVRQASGKGLEWVAEIRSQSNNFATHYAESVKGRVIISRDDSKNTAYLQMNSLRTEDTAVYYCTRRWNNFWGQGTTVTVSSHumanised RKA ′5 (SEQ ID NO: 38)DIQMTQSPSSVSASVGDRVTITCKASENVGTYVSWYQQKPGTAPKLLIYGASNRFTGVPSRFSGSGSATDFTLTINNLQPEDFATYYCGQSYTFPYTFGQGTKVEIK Humanised RKB ′5(SEQ ID NO: 39)DIQMTQSPSSVSASVGDRVTITCKASENVGTYVSWYQQKPGTAPKLLIYGASNRFTGVPSRFSGSGSATDFTLTINNLQPEDFATYYCGQSYTFPYTFGQGTKVEIK Humanised RKC ′5(SEQ ID NO: 40)DIQMTQSPSSVSASVGDRVTITCKASENVGTYVSWYQQKPGTAPKMLIYGASNRFTGVPSRFSGSGSATDFTLTINNLQPEDFATYYCGQSYTFPYTFGQGTKVEIK Humanised RKD ′5(SEQ ID NO: 41)DIQMTQSPSSVSASVGDRVTITCKASENVGTYVSWYQQKPGTAPKMLIYGASNRFTGVPSRFSGSGSATDFTLTINNLQPEDFATYYCGQSYTFPYTFGQGTKVEIK Humanised RKE ′5(SEQ ID NO: 42)NIVMTQSPSSVSASVGDRVTITCKASENVGTYVSWYQQKPGTAPKLLIYGASNRFTGVPDRFTGSGSATDFILTINNLQPEDFATYYCGQSYTFPYTFGQGTKVEIK Humanised RKF ′5(SEQ ID NO: 43)NIVMTQSPSSVSASVGDRVTITCKASENVGTYVSWYQQKPGTAPKMLIYGASNRFTGVPSRFSGSGSATDFILTINNLQPEDFATYYCGQSYTFPYTFGQGTKVEIK Humanised RKG ′5(SEQ ID NO: 44)NIVMTQSPSSVSASVGDRVTITCKASENVGTYVSWYQQKPGTAPKMLIYGASNRFTGVPDRFTGSGSATDFTLTINNLQPEDFATYYCGQSYTFPYTFGQGTKVEIK

The parent antibody may also be a fusion protein comprising analbumin-binding peptide (ABP) sequence (Dennis et al. (2002) “AlbuminBinding As A General Strategy For Improving The Pharmacokinetics OfProteins” J Biol Chem. 277:35035-35043; WO 01/45746). Antibodies of theinvention include fusion proteins with ABP sequences taught by: (i)Dennis et al (2002) J Biol Chem. 277:35035-35043 at Tables III and IV,page 35038; (ii) US 2004/0001827 at [0076]; and (iii) WO 01/45746 atpages 12-13, and all of which are incorporated herein by reference.

In one embodiment, the antibody has been raised to target specific thetumour related antigen α_(v)β₆.

The cell binding agent may be labelled, for example to aid detection orpurification of the agent either prior to incorporation as a conjugate,or as part of the conjugate. The label may be a biotin label. In anotherembodiment, the cell binding agent may be labelled with a radioisotope.

Connection of Linker Unit to Ligand Unit

The Ligand unit is connected to the Linker unit. In one embodiment, theLigand unit is connected to A, where present, of the Linker unit.

In one embodiment, the connection between the Ligand unit and the Linkerunit is through a thioether bond.

In one embodiment, the connection between the Ligand unit and the Linkerunit is through a disulfide bond.

In one embodiment, the connection between the Ligand unit and the Linkerunit is through an amide bond.

In one embodiment, the connection between the Ligand unit and the Linkerunit is through an ester bond.

In one embodiment, the connection between the Ligand unit and the Linkeris formed between a thiol group of a cysteine residue of the Ligand unitand a maleimide group of the Linker unit.

The cysteine residues of the Ligand unit may be available for reactionwith the functional group of the Linker unit to form a connection. Inother embodiments, for example where the Ligand unit is an antibody, thethiol groups of the antibody may participate in interchain disulfidebonds. These interchain bonds may be converted to free thiol groups bye.g. treatment of the antibody with DTT prior to reaction with thefunctional group of the Linker unit.

In some embodiments, the cysteine residue is an introduced into theheavy or light chain of an antibody. Positions for cysteine insertion bysubstitution in antibody heavy or light chains include those describedin Published U.S. Application No. 2007-0092940 and International PatentPublication WO2008070593, which are incorporated herein.

Methods of Treatment

The compounds of the present invention may be used in a method oftherapy. Also provided is a method of treatment, comprisingadministering to a subject in need of treatment atherapeutically-effective amount of a compound of formula I. The term“therapeutically effective amount” is an amount sufficient to showbenefit to a patient. Such benefit may be at least amelioration of atleast one symptom. The actual amount administered, and rate andtime-course of administration, will depend on the nature and severity ofwhat is being treated. Prescription of treatment, e.g. decisions ondosage, is within the responsibility of general practitioners and othermedical doctors.

A compound may be administered alone or in combination with othertreatments, either simultaneously or sequentially dependent upon thecondition to be treated. Examples of treatments and therapies include,but are not limited to, chemotherapy (the administration of activeagents, including, e.g. drugs; surgery; and radiation therapy.

Pharmaceutical compositions according to the present invention, and foruse in accordance with the present invention, may comprise, in additionto the active ingredient, i.e. a compound of formula I, apharmaceutically acceptable excipient, carrier, buffer, stabiliser orother materials well known to those skilled in the art. Such materialsshould be non-toxic and should not interfere with the efficacy of theactive ingredient. The precise nature of the carrier or other materialwill depend on the route of administration, which may be oral, or byinjection, e.g. cutaneous, subcutaneous, or intravenous.

Pharmaceutical compositions for oral administration may be in tablet,capsule, powder or liquid form. A tablet may comprise a solid carrier oran adjuvant. Liquid pharmaceutical compositions generally comprise aliquid carrier such as water, petroleum, animal or vegetable oils,mineral oil or synthetic oil. Physiological saline solution, dextrose orother saccharide solution or glycols such as ethylene glycol, propyleneglycol or polyethylene glycol may be included. A capsule may comprise asolid carrier such a gelatin.

For intravenous, cutaneous or subcutaneous injection, or injection atthe site of affliction, the active ingredient will be in the form of aparenterally acceptable aqueous solution which is pyrogen-free and hassuitable pH, isotonicity and stability. Those of relevant skill in theart are well able to prepare suitable solutions using, for example,isotonic vehicles such as Sodium Chloride Injection, Ringer's Injection,Lactated Ringer's Injection. Preservatives, stabilisers, buffers,antioxidants and/or other additives may be included, as required.

The Compounds and Conjugates can be used to treat proliferative diseaseand autoimmune disease. The term “proliferative disease” pertains to anunwanted or uncontrolled cellular proliferation of excessive or abnormalcells which is undesired, such as, neoplastic or hyperplastic growth,whether in vitro or in vivo.

Examples of proliferative conditions include, but are not limited to,benign, pre-malignant, and malignant cellular proliferation, includingbut not limited to, neoplasms and tumours (e.g., histocytoma, glioma,astrocyoma, osteoma), cancers (e.g. lung cancer, small cell lung cancer,gastrointestinal cancer, bowel cancer, colon cancer, breast carinoma,ovarian carcinoma, prostate cancer, testicular cancer, liver cancer,kidney cancer, bladder cancer, pancreatic cancer, brain cancer, sarcoma,osteosarcoma, Kaposi's sarcoma, melanoma), leukemias, psoriasis, bonediseases, fibroproliferative disorders (e.g. of connective tissues), andatherosclerosis. Other cancers of interest include, but are not limitedto, haematological; malignancies such as leukemias and lymphomas, suchas non-Hodgkin lymphoma, and subtypes such as DLBCL, marginal zone,mantle zone, and follicular, Hodgkin lymphoma, AML, and other cancers ofB or T cell origin.

Examples of autoimmune disease include the following: rheumatoidarthritis, autoimmune demyelinative diseases (e.g., multiple sclerosis,allergic encephalomyelitis), psoriatic arthritis, endocrineophthalmopathy, uveoretinitis, systemic lupus erythematosus, myastheniagravis, Graves' disease, glomerulonephritis, autoimmune hepatologicaldisorder, inflammatory bowel disease (e.g., Crohn's disease),anaphylaxis, allergic reaction, Sjögren's syndrome, type I diabetesmellitus, primary biliary cirrhosis, Wegener's granulomatosis,fibromyalgia, polymyositis, dermatomyositis, multiple endocrine failure,Schmidt's syndrome, autoimmune uveitis, Addison's disease, adrenalitis,thyroiditis, Hashimoto's thyroiditis, autoimmune thyroid disease,pernicious anemia, gastric atrophy, chronic hepatitis, lupoid hepatitis,atherosclerosis, subacute cutaneous lupus erythematosus,hypoparathyroidism, Dressler's syndrome, autoimmune thrombocytopenia,idiopathic thrombocytopenic purpura, hemolytic anemia, pemphigusvulgaris, pemphigus, dermatitis herpetiformis, alopecia arcata,pemphigoid, scleroderma, progressive systemic sclerosis, CREST syndrome(calcinosis, Raynaud's phenomenon, esophageal dysmotility,sclerodactyly, and telangiectasia), male and female autoimmuneinfertility, ankylosing spondolytis, ulcerative colitis, mixedconnective tissue disease, polyarteritis nedosa, systemic necrotizingvasculitis, atopic dermatitis, atopic rhinitis, Goodpasture's syndrome,Chagas' disease, sarcoidosis, rheumatic fever, asthma, recurrentabortion, anti-phospholipid syndrome, farmer's lung, erythemamultiforme, post cardiotomy syndrome, Cushing's syndrome, autoimmunechronic active hepatitis, bird-fancier's lung, toxic epidermalnecrolysis, Alport's syndrome, alveolitis, allergic alveolitis,fibrosing alveolitis, interstitial lung disease, erythema nodosum,pyoderma gangrenosum, transfusion reaction, Takayasu's arteritis,polymyalgia rheumatica, temporal arteritis, schistosomiasis, giant cellarteritis, ascariasis, aspergillosis, Sampter's syndrome, eczema,lymphomatoid granulomatosis, Behcet's disease, Caplan's syndrome,Kawasaki's disease, dengue, encephalomyelitis, endocarditis,endomyocardial fibrosis, endophthalmitis, erythema elevatum et diutinum,psoriasis, erythroblastosis fetalis, eosinophilic faciitis, Shulman'ssyndrome, Felty's syndrome, filariasis, cyclitis, chronic cyclitis,heterochronic cyclitis, Fuch's cyclitis, IgA nephropathy,Henoch-Schonlein purpura, graft versus host disease, transplantationrejection, cardiomyopathy, Eaton-Lambert syndrome, relapsingpolychondritis, cryoglobulinemia, Waldenstrom's macroglobulemia, Evan'ssyndrome, and autoimmune gonadal failure.

In some embodiments, the autoimmune disease is a disorder of Blymphocytes (e.g., systemic lupus erythematosus, Goodpasture's syndrome,rheumatoid arthritis, and type I diabetes), Th1-lymphocytes (e.g.,rheumatoid arthritis, multiple sclerosis, psoriasis, Sjögren's syndrome,Hashimoto's thyroiditis, Graves' disease, primary biliary cirrhosis,Wegener's granulomatosis, tuberculosis, or graft versus host disease),or Th2-lymphocytes (e.g., atopic dermatitis, systemic lupuserythematosus, atopic asthma, rhinoconjunctivitis, allergic rhinitis,Omenn's syndrome, systemic sclerosis, or chronic graft versus hostdisease). Generally, disorders involving dendritic cells involvedisorders of Th1-lymphocytes or Th2-lymphocytes. In some embodiments,the autoimmunie disorder is a T cell-mediated immunological disorder.

In some embodiments, the amount of the Conjugate administered rangesfrom about 0.01 to about 10 mg/kg per dose. In some embodiments, theamount of the Conjugate administered ranges from about 0.01 to about 5mg/kg per dose. In some embodiments, the amount of the Conjugateadministered ranges from about 0.05 to about 5 mg/kg per dose. In someembodiments, the amount of the Conjugate administered ranges from about0.1 to about 5 mg/kg per dose. In some embodiments, the amount of theConjugate administered ranges from about 0.1 to about 4 mg/kg per dose.In some embodiments, the amount of the Conjugate administered rangesfrom about 0.05 to about 3 mg/kg per dose. In some embodiments, theamount of the Conjugate administered ranges from about 0.1 to about 3mg/kg per dose. In some embodiments, the amount of the Conjugateadministered ranges from about 0.1 to about 2 mg/kg per dose.

Drug Loading

The drug loading is the average number of PBD drugs per cell bindingagent, e.g. antibody. Where the compounds of the invention are bound tocysteines, drug loading may range from 1 to 8 drugs (D) per cell bindingagent, i.e. where 1, 2, 3, 4, 5, 6, 7, and 8 drug moieties arecovalently attached to the cell binding agent. Compositions ofconjugates include collections of cell binding agents, e.g. antibodies,conjugated with a range of drugs, from 1 to 8. Where the compounds ofthe invention are bound to lysines, drug loading may range from 1 to 80drugs (D) per cell binding agent, although an upper limit of 40, 20, 10or 8 may be preferred. Compositions of conjugates include collections ofcell binding agents, e.g. antibodies, conjugated with a range of drugs,from 1 to 80, 1 to 40, 1 to 20, 1 to 10 or 1 to 8.

The average number of drugs per antibody in preparations of ADC fromconjugation reactions may be characterized by conventional means such asUV, reverse phase HPLC, HIC, mass spectroscopy, ELISA assay, andelectrophoresis. The quantitative distribution of ADC in terms of p mayalso be determined. By ELISA, the averaged value of p in a particularpreparation of ADC may be determined (Hamblett et al (2004) Clin. CancerRes. 10:7063-7070; Sanderson et al (2005) Clin. Cancer Res. 11:843-852).However, the distribution of p (drug) values is not discernible by theantibody-antigen binding and detection limitation of ELISA. Also, ELISAassay for detection of antibody-drug conjugates does not determine wherethe drug moieties are attached to the antibody, such as the heavy chainor light chain fragments, or the particular amino acid residues. In someinstances, separation, purification, and characterization of homogeneousADC where p is a certain value from ADC with other drug loadings may beachieved by means such as reverse phase HPLC or electrophoresis. Suchtechniques are also applicable to other types of conjugates.

For some antibody-drug conjugates, p may be limited by the number ofattachment sites on the antibody. For example, an antibody may have onlyone or several cysteine thiol groups, or may have only one or severalsufficiently reactive thiol groups through which a linker may beattached. Higher drug loading, e.g. p>5, may cause aggregation,insolubility, toxicity, or loss of cellular permeability of certainantibody-drug conjugates.

Typically, fewer than the theoretical maximum of drug moieties areconjugated to an antibody during a conjugation reaction. An antibody maycontain, for example, many lysine residues that do not react with thedrug-linker intermediate (D-L) or linker reagent. Only the most reactivelysine groups may react with an amine-reactive linker reagent. Also,only the most reactive cysteine thiol groups may react with athiol-reactive linker reagent. Generally, antibodies do not containmany, if any, free and reactive cysteine thiol groups which may belinked to a drug moiety. Most cysteine thiol residues in the antibodiesof the compounds exist as disulfide bridges and must be reduced with areducing agent such as dithiothreitol (DTT) or TCEP, under partial ortotal reducing conditions. The loading (drug/antibody ratio) of an ADCmay be controlled in several different manners, including: (i) limitingthe molar excess of drug-linker intermediate (D-L) or linker reagentrelative to antibody, (ii) limiting the conjugation reaction time ortemperature, and (iii) partial or limiting reductive conditions forcysteine thiol modification.

Certain antibodies have reducible interchain disulfides, i.e. cysteinebridges. Antibodies may be made reactive for conjugation with linkerreagents by treatment with a reducing agent such as DTT(dithiothreitol). Each cysteine bridge will thus form, theoretically,two reactive thiol nucleophiles. Additional nucleophilic groups can beintroduced into antibodies through the reaction of lysines with2-iminothiolane (Traut's reagent) resulting in conversion of an amineinto a thiol. Reactive thiol groups may be introduced into the antibody(or fragment thereof) by engineering one, two, three, four, or morecysteine residues (e.g., preparing mutant antibodies comprising one ormore non-native cysteine amino acid residues). U.S. Pat. No. 7,521,541teaches engineering antibodies by introduction of reactive cysteineamino acids.

Cysteine amino acids may be engineered at reactive sites in an antibodyand which do not form intrachain or intermolecular disulfide linkages(Junutula, et al., 2008b Nature Biotech., 26(8):925-932; Dornan et al(2009) Blood 114(13):2721-2729; U.S. Pat. Nos. 7,521,541; 7,723,485;WO2009/052249). The engineered cysteine thiols may react with linkerreagents or the drug-linker reagents of the present invention which havethiol-reactive, electrophilic groups such as maleimide or alpha-haloamides to form ADC with cysteine engineered antibodies and the PBD drugmoieties. The location of the drug moiety can thus be designed,controlled, and known. The drug loading can be controlled since theengineered cysteine thiol groups typically react with thiol-reactivelinker reagents or drug-linker reagents in high yield. Engineering anIgG antibody to introduce a cysteine amino acid by substitution at asingle site on the heavy or light chain gives two new cysteines on thesymmetrical antibody. A drug loading near 2 can be achieved with nearhomogeneity of the conjugation product ADC.

Where more than one nucleophilic or electrophilic group of the antibodyreacts with a drug-linker intermediate, or linker reagent followed bydrug moiety reagent, then the resulting product is a mixture of ADCcompounds with a distribution of drug moieties attached to an antibody,e.g. 1, 2, 3, etc. Liquid chromatography methods such as polymericreverse phase (PLRP) and hydrophobic interaction (HIC) may separatecompounds in the mixture by drug loading value. Preparations of ADC witha single drug loading value (p) may be isolated, however, these singleloading value ADCs may still be heterogeneous mixtures because the drugmoieties may be attached, via the linker, at different sites on theantibody.

Thus the antibody-drug conjugate compositions of the invention includemixtures of antibody-drug conjugate compounds where the antibody has oneor more PBD drug moieties and where the drug moieties may be attached tothe antibody at various amino acid residues.

In one embodiment, the average number of dimer pyrrolobenzodiazepinegroups per cell binding agent is in the range 1 to 20. In someembodiments the range is selected from 1 to 8, 2 to 8, 2 to 6, 2 to 4,and 4 to 8.

In some embodiments, there is one dimer pyrrolobenzodiazepine group percell binding agent.

Includes Other Forms

Unless otherwise specified, included in the above are the well knownionic, salt, solvate, and protected forms of these substituents. Forexample, a reference to carboxylic acid (—COOH) also includes theanionic (carboxylate) form (—COO⁻), a salt or solvate thereof, as wellas conventional protected forms. Similarly, a reference to an aminogroup includes the protonated form (—N⁺HR¹R²), a salt or solvate of theamino group, for example, a hydrochloride salt, as well as conventionalprotected forms of an amino group. Similarly, a reference to a hydroxylgroup also includes the anionic form (—O⁻), a salt or solvate thereof,as well as conventional protected forms.

Salts

It may be convenient or desirable to prepare, purify, and/or handle acorresponding salt of the active compound, for example, apharmaceutically-acceptable salt. Examples of pharmaceuticallyacceptable salts are discussed in Berge, et al., J. Pharm. Sci., 66,1-19 (1977).

For example, if the compound is anionic, or has a functional group whichmay be anionic (e.g. —COOH may be —COO), then a salt may be formed witha suitable cation. Examples of suitable inorganic cations include, butare not limited to, alkali metal ions such as Na⁺ and K⁺, alkaline earthcations such as Ca²⁺ and Mg²⁺, and other cations such as Al⁺³. Examplesof suitable organic cations include, but are not limited to, ammoniumion (i.e. NH₄ ⁺) and substituted ammonium ions (e.g. NH₃R⁺, NH₂R₂ ⁺,NHR₃ ⁺, NR₄ ⁺). Examples of some suitable substituted ammonium ions arethose derived from: ethylamine, diethylamine, dicyclohexylamine,triethylamine, butylamine, ethylenediamine, ethanolamine,diethanolamine, piperazine, benzylamine, phenylbenzylamine, choline,meglumine, and tromethamine, as well as amino acids, such as lysine andarginine. An example of a common quaternary ammonium ion is N(CH₃)₄ ⁺.

If the compound is cationic, or has a functional group which may becationic (e.g. —NH₂ may be —NH₃ ⁺), then a salt may be formed with asuitable anion. Examples of suitable inorganic anions include, but arenot limited to, those derived from the following inorganic acids:hydrochloric, hydrobromic, hydroiodic, sulfuric, sulfurous, nitric,nitrous, phosphoric, and phosphorous.

Examples of suitable organic anions include, but are not limited to,those derived from the following organic acids: 2-acetyoxybenzoic,acetic, ascorbic, aspartic, benzoic, camphorsulfonic, cinnamic, citric,edetic, ethanedisulfonic, ethanesulfonic, fumaric, glucheptonic,gluconic, glutamic, glycolic, hydroxymaleic, hydroxynaphthalenecarboxylic, isethionic, lactic, lactobionic, lauric, maleic, malic,methanesulfonic, mucic, oleic, oxalic, palmitic, pamoic, pantothenic,phenylacetic, phenylsulfonic, propionic, pyruvic, salicylic, stearic,succinic, sulfanilic, tartaric, toluenesulfonic, and valeric. Examplesof suitable polymeric organic anions include, but are not limited to,those derived from the following polymeric acids: tannic acid,carboxymethyl cellulose.

Solvates

It may be convenient or desirable to prepare, purify, and/or handle acorresponding solvate of the active compound. The term “solvate” is usedherein in the conventional sense to refer to a complex of solute (e.g.active compound, salt of active compound) and solvent. If the solvent iswater, the solvate may be conveniently referred to as a hydrate, forexample, a mono-hydrate, a di-hydrate, a tri-hydrate, etc.

Carbinolamines

The invention includes compounds where a solvent adds across the iminebond of the PBD moiety, which is illustrated below where the solvent iswater or an alcohol (R^(A)OH, where R_(A) is C₁₋₄ alkyl):

These forms can be called the carbinolamine and carbinolamine etherforms of the PBD. The balance of these equilibria depend on theconditions in which the compounds are found, as well as the nature ofthe moiety itself.

These particular compounds may be isolated in solid form, for example,by lyophilisation.

Isomers

Certain compounds may exist in one or more particular geometric,optical, enantiomeric, diasteriomeric, epimeric, atropic,stereoisomeric, tautomeric, conformational, or anomeric forms, includingbut not limited to, cis- and trans-forms; E- and Z-forms; c-, t-, and r-forms; endo- and exo-forms; R-, S-, and meso-forms; D- and L-forms; d-and l-forms; (+) and (−) forms; keto-, enol-, and enolate-forms; syn-and anti-forms; synclinal- and anticlinal-forms; α- and β-forms; axialand equatorial forms; boat-, chair-, twist-, envelope-, andhalfchair-forms; and combinations thereof, hereinafter collectivelyreferred to as “isomers” (or “isomeric forms”).

Note that, except as discussed below for tautomeric forms, specificallyexcluded from the term “isomers”, as used herein, are structural (orconstitutional) isomers (i.e. isomers which differ in the connectionsbetween atoms rather than merely by the position of atoms in space). Forexample, a reference to a methoxy group, —OCH₃, is not to be construedas a reference to its structural isomer, a hydroxymethyl group, —CH₂OH.Similarly, a reference to ortho-chlorophenyl is not to be construed as areference to its structural isomer, meta-chlorophenyl. However, areference to a class of structures may well include structurallyisomeric forms falling within that class (e.g. alkyl includes n-propyland iso-propyl; butyl includes n-, iso-, sec-, and tert-butyl;methoxyphenyl includes ortho-, meta-, and para-methoxyphenyl).

The above exclusion does not pertain to tautomeric forms, for example,keto-, enol-, and enolate-forms, as in, for example, the followingtautomeric pairs: keto/enol (illustrated below), imine/enamine,amide/imino alcohol, amidine/amidine, nitroso/oxime,thioketone/enethiol, N-nitroso/hyroxyazo, and nitro/aci-nitro.

Note that specifically included in the term “isomer” are compounds withone or more isotopic substitutions. For example, H may be in anyisotopic form, including ¹H, ²H (D), and ³H (T); C may be in anyisotopic form, including ¹³C, and ¹⁴C; O may be in any isotopic form,including ¹⁶O and ¹⁸O; and the like.

Unless otherwise specified, a reference to a particular compoundincludes all such isomeric forms, including (wholly or partially)racemic and other mixtures thereof. Methods for the preparation (e.g.asymmetric synthesis) and separation (e.g. fractional crystallisationand chromatographic means) of such isomeric forms are either known inthe art or are readily obtained by adapting the methods taught herein,or known methods, in a known manner.

General Synthetic Routes

The synthesis of PBD compounds is extensively discussed in the followingreferences, which discussions are incorporated herein by reference:

a) WO 00/12508 (pages 14 to 30);

b) WO 2005/023814 (pages 3 to 10);

c) WO 2004/043963 (pages 28 to 29); and

d) WO 2005/085251 (pages 30 to 39).

Synthesis Route

The compounds of the present invention, where R¹⁰ and R¹¹ form anitrogen-carbon double bond between the nitrogen and carbon atoms towhich they are bound, can be synthesised from a compound of Formula 2:

where R², R⁶, R⁷, R⁹, R^(6′), R^(7′), R^(9′), R¹², X, X′ and R″ are asdefined for compounds of formula I, Prot^(N) is a nitrogen protectinggroup for synthesis and Prot^(O) is a protected oxygen group forsynthesis or an oxo group, by deprotecting the imine bond by standardmethods.

The compound produced may be in its carbinolamine or carbinolamine etherform depending on the solvents used. For example if Prot^(N) is Allocand Prot^(O) is an oxygen protecting group for synthesis, then thedeprotection is carried using palladium to remove the N10 protectinggroup, followed by the elimination of the oxygen protecting group forsynthesis. If Prot^(N) is Troc and Prot^(O) is an oxygen protectinggroup for synthesis, then the deprotection is carried out using a Cd/Pbcouple to yield the compound of formula (I). If Prot^(N) is SEM, or ananalogous group, and Prot^(O) is an an oxo group, then the oxo group canbe removed by reduction, which leads to a protected carbinolamineintermediate, which can then be treated to remove the SEM protectinggroup, followed by the elimination of water. The reduction of thecompound of Formula 2 can be accomplished by, for example, lithiumtetraborohydride, whilst a suitable means for removing the SEMprotecting group is treatment with silica gel.

Compounds of formula 2 can be synthesised from a compound of formula 3:

where R¹², R⁶, R⁷, R⁹, R^(6′), R^(7′), R^(9′), X, X′ and R″ are asdefined for compounds of formula 2, by coupling an organometallicderivative comprising R², such as an organoboron derivative. Theorganoboron derivative may be a boronate or boronic acid.

The couplings described above are usually carried out in the presence ofa palladium catalyst, for example Pd(PPh₃)₄, Pd(OCOCH₃)₂, PdCl₂,Pd₂(dba)₃. The coupling may be carried out under standard conditions, ormay also be carried out under microwave conditions.

Compounds of formula I where R¹⁰ and R^(10′) are H and R¹¹ and R^(11′)are SO_(z)M, can be synthesised from compounds of formula I where R¹⁰and R¹¹ form a nitrogen-carbon double bond between the nitrogen andcarbon atoms to which they are bound, by the addition of the appropriatebisulphite salt or sulphinate salt, followed by an appropriatepurification step. Further methods are described in GB 2 053 894, whichis herein incorporated by reference.

The PBD monomer components can be synthesised and linked in a similarmanner to that described in WO 2005/085259, where one monomer unit islinked to the tether unit, following by reaction with a second monomerunit.

Nitrogen Protecting Groups for Synthesis

Nitrogen protecting groups for synthesis are well known in the art. Inthe present invention, the protecting groups of particular interest arecarbamate nitrogen protecting groups and hemi-aminal nitrogen protectinggroups.

Carbamate nitrogen protecting groups have the following structure:

wherein R′¹⁰ is R as defined above. A large number of suitable groupsare described on pages 503 to 549 of Greene, T. W. and Wuts, G.M.,Protective Groups in Organic Synthesis, 3^(rd) Edition, John Wiley &Sons, Inc., 1999, which is incorporated herein by reference.

Particularly preferred protecting groups include Troc, Teoc, Fmoc, BOC,Doc, Hoc, TcBOC, 1-Adoc and 2-Adoc.

Other possible groups are nitrobenzyloxycarbonyl (e.g.4-nitrobenzyloxycarbonyl) and 2-(phenylsulphonyl)ethoxycarbonyl.

Those protecting groups which can be removed with palladium catalysisare not preferred, e.g. Alloc.

Hemi-aminal nitrogen protecting groups have the following structure:

wherein R′¹⁰ is R as defined above. A large number of suitable groupsare described on pages 633 to 647 as amide protecting groups of Greene,T. W. and Wuts, G.M., Protective Groups in Organic Synthesis, 3rdEdition, John Wiley & Sons, Inc., 1999, which is incorporated herein byreference. The groups disclosed herein can be applied to compounds ofthe present invention. Such groups include, but are not limited to, SEM,MOM, MTM, MEM, BOM, nitro or methoxy substituted BOM, Cl₃CCH₂OCH₂—.

Protected Oxygen Group for Synthesis

Protected oxygen group for synthesis are well known in the art. A largenumber of suitable oxygen protecting groups are described on pages 23 to200 of Greene, T. W. and Wuts, G.M., Protective Groups in OrganicSynthesis, 3^(rd) Edition, John Wiley & Sons, Inc., 1999, which isincorporated herein by reference.

Classes of particular interest include silyl ethers, methyl ethers,alkyl ethers, benzyl ethers, esters, acetates, benzoates, carbonates,and sulfonates.

Preferred oxygen protecting groups include acetates, TBS and THP.

Synthesis of Drug Conjugates

Conjugates can be prepared as previously described. Linkers having amaleimidyl group (A), a peptide group (L¹) and self-immolative group(L²) can be prepared as described in U.S. Pat. No. 6,214,345. Linkershaving a maleimidyl group (A) and a peptide group (L¹) can be preparedas described in WO 2009-0117531. Other linkers can be prepared accordingto the references cited herein or as known to the skilled artisan.

Linker-Drug compounds can be prepared according to methods known in theart. Linkage of amine-based X substituents (of the PDB dimer Drug unit)to active groups of the Linker units can be performed according tomethods generally described in U.S. Pat. Nos. 6,214,345 and 7,498,298;and WO 2009-0117531, or as otherwise known to the skilled artisan.

Antibodies can be conjugated to Linker-Drug compounds as described inDoronina et al., Nature Biotechnology, 2003, 21, 778-784). Briefly,antibodies (4-5 mg/mL) in PBS containing 50 mM sodium borate at pH 7.4are reduced with tris(carboxyethyl)phosphine hydrochloride (TCEP) at 37°C. The progress of the reaction, which reduces interchain disulfides, ismonitored by reaction with 5,5′-dithiobis(2-nitrobenzoic acid) andallowed to proceed until the desired level of thiols/mAb is achieved.The reduced antibody is then cooled to 0° C. and alkylated with 1.5equivalents of maleimide drug-linker per antibody thiol. After 1 hour,the reaction is quenched by the addition of 5 equivalents of N-acetylcysteine. Quenched drug-linker is removed by gel filtration over a PD-10column. The ADC is then sterile-filtered through a 0.22 pm syringefilter. Protein concentration can be determined by spectral analysis at280 nm and 329 nm, respectively, with correction for the contribution ofdrug absorbance at 280 nm. Size exclusion chromatography can be used todetermine the extent of antibody aggregation, and RP-HPLC can be used todetermine the levels of remaining NAC-quenched drug-linker.

Further Preferences

The following preferences may apply to all aspects of the invention asdescribed above, or may relate to a single aspect. The preferences maybe combined together in any combination.

In some embodiments, R^(6′), R^(7′), R^(9′), R^(10′), R^(11′), and Y′are preferably the same as R⁶, R⁷, R⁹, R¹⁰, R¹¹ and Y respectively.

Dimer Link

Y and Y′ are preferably 0.

R″ is preferably a C₃₋₇ alkylene group with no substituents. Morepreferably R″ is a C₃, C₅ or C₇ alkylene. Most preferably, R″ is a C₃ orC₅ alkylene.

R⁶ to R⁹

R⁹ is preferably H.

R⁶ is preferably selected from H, OH, OR, SH, NH₂, nitro and halo, andis more preferably H or halo, and most preferably is H.

R⁷ is preferably selected from H, OH, OR, SH, SR, NH₂, NHR, NRR′, andhalo, and more preferably independently selected from H, OH and OR,where R is preferably selected from optionally substituted C₁₋₇ alkyl,C₃₋₁₀ heterocyclyl and C₅₋₁₀ aryl groups. R may be more preferably aC₁₋₄ alkyl group, which may or may not be substituted. A substituent ofinterest is a C₅₋₆ aryl group (e.g. phenyl). Particularly preferredsubstituents at the 7-positions are OMe and OCH₂Ph. Other substituentsof particular interest are dimethylamino (i.e. —NMe₂); —(OC₂H₄)_(q)OMe,where q is from 0 to 2; nitrogen-containing C₆ heterocyclyls, includingmorpholino, piperidinyl and N-methyl-piperazinyl.

These preferences apply to R^(9′), R^(6′) and R^(7′) respectively.

R²

A in R² may be phenyl group or a 05.7 heteroaryl group, for examplefuranyl, thiophenyl and pyridyl. In some embodiments, A is preferablyphenyl.

X is a group selected from the list comprising: OH, SH, CO₂H, COH,N═C═O, NHNH₂, CONHNH₂,

and NHR^(N), wherein R^(N) is selected from the group comprising H andC₁₋₄ alkyl. X may preferably be: OH, SH, CO₂H, —N═C═O or NHR^(N), andmay more preferably be: OH, SH, CO₂H, —N═C═O or NH₂. Particularlypreferred groups include: OH, SH and NH₂, with NH₂ being the mostpreferred group.

Q²—X may be on any of the available ring atoms of the C₅₋₇ aryl group,but is preferably on a ring atom that is not adjacent the bond to theremainder of the compound, i.e. it is preferably β or γ to the bond tothe remainder of the compound. Therefore, where the C₅₋₇ aryl group (A)is phenyl, the substituent (Q²—X) is preferably in the meta- orpara-positions, and more preferably is in the para-position.

In some embodiments, Q¹ is a single bond. In these embodiments, Q² isselected from a single bond and —Z—(CH₂)_(n)—, where Z is selected froma single bond, O, S and NH and is from 1 to 3. In some of theseembodiments, Q² is a single bond. In other embodiments, Q² is—Z—(CH₂)_(n)—. In these embodiments, Z may be O or S and n may be 1 or nmay be 2. In other of these embodiments, Z may be a single bond and nmay be 1.

In other embodiments, Q¹ is —CH═CH—.

In some embodiments, R² may be -A-CH₂—X and -A-X. In these embodiments,X may be OH, SH, CO₂H, COH and NH₂. In particularly preferredembodiments, X may be NH₂.

R¹²

In one embodiment, R¹² is ═CH₂.

In one embodiment, R¹² is ═CH—R^(D1). Within the PBD compound, the group═CH—R^(D1) may have either configuration shown below:

In one embodiment, the configuration is configuration (I).

R^(D1) and R^(D2) (when present) may be selected from R, and inparticular from C₁₋₃ alkyl, e.g. methyl, ethyl, propyl. In some of theseembodiments, R^(D1) and R^(D2) (when present) are methyl.

R¹⁰ and R¹¹

In one embodiment, R¹⁰ is H, and R¹¹ is OH, OR^(A), where R^(A) is C₁₋₄alkyl. In some of these embodiments, R^(A) is methyl.

In another embodiment, R¹⁰ and R¹¹ form a nitrogen-carbon double bondbetween the nitrogen and carbon atoms to which they are bound.

In another embodiment, R¹⁰ is H and R¹¹ is SO_(z)M, where z is 2 or 3and M is a monovalent pharmaceutically acceptable cation.

M and z

It is preferred that M and M′ are monovalent pharmaceutically acceptablecations, and are more preferably Na⁺.

z is preferably 3.

Particularly preferred compounds of the present invention are of formulaIa:

where R^(1a) is selected from Me and Ph, and the amino group is ateither the meta or para positions of the phenyl group.

3^(rd) Aspect

The preferences expressed above for the first aspect may apply to thecompounds of this aspect, where appropriate.

When R¹⁰ is carbamate nitrogen protecting group, it may preferably beTeoc, Fmoc and Troc, and may more preferably be Troc.

When R¹¹ is O-Prot^(O), wherein Prot^(O) is an oxygen protecting group,Prot^(O) may preferably be TBS or THP, and may more preferably be TBS.

When R¹⁰ is a hemi-aminal nitrogen protecting group, it may preferablybe MOM, BOM or SEM, and may more preferably be SEM.

The preferences for compounds of formula I apply as appropriate to D inthe sixth aspect of the invention.

EXAMPLES

General Experimental Methods

Reaction progress was monitored by thin-layer chromatography (TLC) usingMerck Kieselgel 60 F254 silica gel, with fluorescent indicator onaluminium plates. Visualisation of TLC was achieved with UV light oriodine vapour unless otherwise stated. Flash chromatography wasperformed using Merck Kieselgel 60 F254 silica gel. Extraction andchromatography solvents were bought and used without furtherpurification from Fisher Scientific, U.K. All chemicals were purchasedfrom Aldrich, Lancaster or BDH.

Method for LS/MS Spectrometer:

LC/MS (Shimazu LCMS-2020) using a mobile phase of water (A) (formic acid0.1%) and acetonitrile (B) (formic acid 0.1%). Gradient: initialcomposition 5% B held over 0.25 min, then increase from 5% B to 100% Bover a 2 min period. The composition was held for 0.50 min at 100% B,then returned to 5% B in 0.05 minutes and hold there for 0.05 min. Totalgradient run time equals 3 min. Flow rate 0.8 mL/min. Wavelengthdetection range: 190 to 800 nm. Oven temperature: 50° C. Column: WatersAcquity UPLC BEH Shield RP18 1.7 μm 2.1×50 mm.

Method for Preparative HPLC:

HPLC (Shimadzu UFLC) was run using a mobile phase of water (0.1% formicacid) A and acetonitrile (0.1% formic acid) B.

Wavelength detection range: 254 nm.

Column: Phenomenex Gemini 5p C18 150×21-20 mm.

Gradient:

B t = 0 13% t = 15.00 95% t = 17.00 95% t = 17.10 13% t = 20.00 13%

Total gradient run time is 20 min; flow rate 20.00 mL/min.

Example 1 (i) tert-Butyl(11S,11aS)-2-(4-(2-((R)-1-(((S)-1-((9H-fluoren-9-yl)methoxy)-4-methyl-1,2-dioxopentan-3-yl)amino)-1-oxopropan-2-yl)hydrazinyl)phenyl)-11-((tert-butyldimethylsilyl)oxy)-8-hydroxy-7-methoxy-5-oxo-11,11a-dihydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-10(5H)-carboxylate(10)

(a)(S)-5-(((tert-butyldimethylsilyl)oxy)methyl)-1-(5-methoxy-2-nitro-4-((triisopropylsilyl)oxy)benzoyl)-4,5-dihydro-1H-pyrrol-3-yltrifluoromethanesulfonate (2)

Ketone monomer 1 (20 g, 34 mmol, 1 eq.) was solubilised in dry CH₂Cl₂(350 mL), before 2,6-lutidine (16.06 mL, 0.137 mol, 4 eq.) was added andthe mixture cooled to −50° C. Triflic anhydride (17.37 mL, 0.1 mol, 3eq) was then added dropwise maintaining the temperature below −40° C.When the reaction was completed, excess Tf₂O was quenched with H₂O (250mL). The layers were separated and the organics were washed further withaqueous NaHCO_(3 (sat.)) (200 mL) and brine (200 mL). The crude mixturewas purified by silica gel chromatography (gradient elution: 100% hexaneto 90:10 v/v hexane/EtOAc) to afford pure product 2 as brown foam(22.065 g, 89% yield). Analytical data: ES⁺=2.39 min, m/z 1448.20[2M+Na]⁺

(b)(S)-(4-(4-aminophenyl)-2-(((tert-butyldimethylsilyl)oxy)methyl)-2,3-dihydro-1H-pyrrol-1-yl)(5-methoxy-2-nitro-4-((triisopropylsilyl)oxy)phenyl)methanone(3)

Pd(PPh₃)₄ (609 mg, 520 mmol, 0.02 eq.) was added to a stirred mixture oftriflate 2 (18.8 g, 26.3 mmol, 1 eq.), 4-aminophenylboronic acid pinacolester (8.64 g, 39.4 mmol, 1.5 eq) and Na₂CO₃ (12.7 g, 120 mmol, 4.6 eq.)in MeOH (75 mL), toluene (150 mL) and water (75 mL). The reactionmixture was allowed to stir at 30° C. under an argon atmosphere for 24hours after which time all the triflate has been consumed. The reactionmixture was then evaporated to dryness before the residue was taken upin EtOAc (400 mL) and washed with H₂O (2×250 mL), brine (250 mL), dried(MgSO₄), filtered and evaporated under reduced pressure to provide thecrude product. Purification by flash chromatography (gradient elution:100% hexane to 80:20 v/v hexane/EtOAc) afforded product 3 as a yellowishfoam (11.058 g, 64% yield). Analytical data: ES⁺=2.20 min, m/z notobserved.

(c) (9H-fluoren-9-yl)methyl(S)-(4-(5-(((tert-butyldimethylsilyl)oxy)methyl)-1-(5-methoxy-2-nitro-4-((triisopropylsilyl)oxy)benzoyl)-4,5-dihydro-1H-pyrrol-3-yl)phenyl)carbamate(4)

To a dry round bottom flask was added aniline 3 (10.05 g, 15.3 mmol),Fmoc-Val-Ala-OH (6.3 g, 15.3 mmol) and dry CH₂Cl₂ (500 mL). The flaskwas then purged 3 times with argon before EEDQ (3.79 mg, 15.3 mmol) wasadded and the mixture left to stir at room temperature. The reaction wasfollowed by LCMS and left to stir for 16 hours. The reaction wasquenched with H₂O (300 mL). The separated organics were washed withbrine (250 mL), dried over MgSO₄, filtered and the solvent removed invacuo. The crude product was purified by silica gel chromatography(gradient elution 80:20 v/v hexane/EtOAc to 50:50 v/v hexane/EtOAc) toafford pure product 4 (13.821 g, 86% yield). Analytical data: ES⁺=2.37min, m/z 1071.65 [M+Na]⁺

(d) (9H-fluoren-9-yl)methyl(S)-(4-(1-(2-amino-5-methoxy-4-((triisopropylsilyl)oxy)benzoyl)-5-(((tert-butyldimethylsilyl)oxy)methyl)-4,5-dihydro-1H-pyrrol-3-yl)phenyl)carbamate(5)

Monomer 4 (16.821 g, 16.04 mmol, 1 eq.) was solubilised in a mixture ofmethanol (600 mL) and formic acid (30 mL). Zinc dust was added slowly(10.49, 160 mmol, 10 eq.) and an exotherm was observed (temperature wentup to 30° C.). After 25 minutes the reaction looked complete by TLC(eluent 1:1 v/v CH₂Cl₂/Et₂O). The reaction mixture was filtered throughcelite, the pad was washed with EtOAc (200 mL) and the solvent removedinto vacuo. The dried crude was taken up in EtOAc (300 mL) and washedwith H₂O (200 mL), NaHCO₃ (200 mL) and brine (200 mL), dried (MgSO₄),filtered and evaporated under reduced pressure. The crude product waspurified by silica gel chromatography (elution 2:1 v/v hexane/EtOAc) toafford pure product 5 (12.642 g, 77% yield). Analytical data: ES⁺=2.36min, m/z 1018.35 [M+H]⁺

(e) (9H-fluoren-9-yl)methyl(S)-(4-(1-(2-((tert-butoxycarbonyl)amino)-5-methoxy-4-((triisopropylsilyl)oxy)benzoyl)-5-(((tert-butyldimethylsilyl)oxy)methyl)-4,5-dihydro-1H-pyrrol-3-yl)phenyl)carbamate (6)

Monomer 5 (14.913 g, 14.6 mmol, 1 eq.), together with Boc anhydride(3.83 g, 17.5 mmol, 1.2 eq.) was heated to 70° C., 25 mL of CH₂Cl₂ werealso added to help solubilising the starting material and wassubsequently left to evaporate with heating. The reaction was completeafter 2 hours. The reaction mixture was put straight away on silica gelcolumn chromatography (gradient elution:100% hexane to 65:35 v/vhexane/EtOAc) to give pure product 6 as a light yellow foam (13.2 g, 80%yield). Analytical data: ES⁺=2.53 min, m/z 1118.35 [M+H]⁺

(t) (9H-fluoren-9-yl)methyl(S)-(4-(1-(2-((tert-butoxycarbonyl)amino)-5-methoxy-4-((triisopropylsilyl)oxy)benzoyl)-5-(hydroxymethyl)-4,5-dihydro-1H-pyrrol-3-yl)phenyl)carbamate(7)

Monomer 6 (13.2 g, 13.9 mmol) was solubilised in a mixture of 7:1:1:2v/v of AcOH/MeOH/THF/H₂O and left to stir for 16 hours. The solventswere subsequently removed and the crude taken up in EtOAc (300 mL),washed with H₂O (200 mL), NaHCO₃ (2×200 mL) and brine (150 mL) beforebeing dried (MgSO₄), filtered and evaporated under reduced pressure. Thecrude product was purified by silica gel chromatography (gradientelution: 100% hexane to 100% EtOAc) to afford pure product 7 (11.168 g,94% yield). Analytical data: ES⁺=2.23 min, m/z not observed.

(g) tert-Butyl(11S,11aS)-2-(4-(2-((R)-1-(((S)-1-(9H-fluoren-9-yl)methoxy)-4-methyl-1,2-dioxopentan-3-yl)amino)-1-oxopropan-2-yl)hydrazinyl)phenyl)-11-hydroxy-7-methoxy-5-oxo-8-((triisopropylsilyl)oxy)-11,11a-dihydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-10(5H)-carboxylate(8)

In a dry flask purged with argon, dry DMSO (1.55 mL, 21.9 mmol, 2.5 eq.)was added to a solution of oxaly chloride (0.89 mL, 10.5 mmol, 1.2eq.)in dry CH₂Cl₂ (50 mL) at −78° C. The mixture was left to stir for 20minutes before monomer 7 (8.8 g, 8.76 mmol, 1 eq.) in CH₂Cl₂ (100 mL)was added. The mixture was left to stir for 1.5 hours at −78° C. beforeTEA (6.11 mL, 43.8 mmol, 5eq.) was added and the mixture left to warm toroom temperature.

After another 2 hours the reaction was quenched with 0.1M aqueous HCl(100 mL). The layers were separated and the organics were washed furtherwith H₂O (150 mL) and brine (150 mL) before being dried (MgSO4),filtered and evaporated under reduced pressure. The crude product waspurified by silica gel chromatography (gradient elution: 100% hexane to1:1:0.1 hexane/EtOAc/MeOH) to afford a 6:4 mixture of product 7 andopened aldehyde monomer. That mixture was stirred overnight in CHCl₃ togive 100% cyclised product 7 (8.8 g, 100% yield). Analytical data:ES⁺=2.19 min, m/z not observed.

(h) tert-Butyl(11S,11aS)-2-(4-(2-((R)-1-(((S)-1-((9H-fluoren-9-yl)methoxy)-4-methyl-1,2-dioxopentan-3-yl)amino)-1-oxopropan-2-yl)hydrazinyl)phenyl)-11-((tert-butyldimethylsilyl)oxy)-7-methoxy-5-oxo-8-((triisopropylsilyl)oxy)-11,11a-dihydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-10(5H)-carboxylate(9)

Monomer 8 (8.8 g, 8.78 mmol, 1 eq.) was solubilised in CH₂Cl₂ (200 mL).The mixture was cooled to 0° C. before slowly adding 2,6-lutidine (4 mL,35.1 mmol, 4 eq.) and TBS-OTf (6 mL, 26.3 mmol, 3 eq.). The mixture wassubsequently left to warm to room temperature.

Once the reaction was complete, the mixture was washed with aqueousNH₄Cl_((sat.)) (150 mL), H₂O (150 mL), aqueous NaHCO_(3 (sat.)) (150 mL)and brine (150 mL) before being dried (MgSO₄), filtered and evaporatedunder reduced pressure. The crude product was purified by silica gelchromatography (gradient elution: 100% hexane to 6:4 v/v hexane/EtOAc)to afford pure product 9 (6.18 g, 70% yield). Analytical data: ES⁺=2.53min, m/z not observed.

(i) tert-Butyl(11S,11aS)-2-(4-(2-((R)-1-(((S)-1-((9H-fluoren-9-yl)methoxy)-4-methyl-1,2-dioxopentan-3-yl)amino)-1-oxopropan-2-yl)hydrazinyl)phenyl)-11-((tert-butyldimethylsilyl)oxy)-8-hydroxy-7-methoxy-5-oxo-11,11a-dihydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-10(5H)-carboxylate(10)

Monomer 9 (3.1 g, 2.84 mmol, 1 eq.) was solubilised in DMF (8 mL) andH₂O (0.2 mL). LiOAc.H₂O (290 mg, 2.84 mmol, 1 eq.) was added. Then asmuch water as possible was added to the mixture without making thestarting material crash out (3.5 mL). Once complete (=1.5 hours) thereaction was quenched by aqueous citric acid (pH=3, 50 mL) and extractedwith EtOAc (3×150 mL). The combined organics were washed with H₂O (150mL) and brine (100 mL) before being dried (MgSO₄), filtered andevaporated under reduced pressure. The crude product was purified bysilica gel chromatography (gradient elution: 6:4 v/v hexane/EtOAc to6:4:1 v/v hexane/EtOAc/MeOH) to afford pure product 10 (2.219 g, 83%yield). Analytical data: ES⁺=1.91 min, m/z 960.55 [M+H]⁺

(ii)1-(3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanamido)-N-((2S)-1-((2S)-1-((4-(7-methoxy-8-((5-((7-methoxy-2-methylene-5-oxo-2,3,5,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-5-oxo-5,11a-dihydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-3,6,9,12,15,18, 21, 24-octaoxaheptacosan-27-amide (15)

Compound 11 is compound 27 in WO 2005/085259.

(a) tert-Butyl(11S,11aS)-8-((5-iodopentyl)oxy)-7-methoxy-2-methylene-5-oxo-11-((tetrahydro-2H-pyran-2-yl)oxy)-2,3,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-10(5H)-carboxylate(12)

Monomer 11 (200 mg, 0.43 mmol, 1 eq.) was solubilised in DMF (4 mL).1,5-Diiodopentate (320 μL, 2.17 mmol, 5eq.) and K2CO₃ (93 mg, 0.75 mmol,1 eq.) were added and the mixture heated to 60° C. Upon completion themixture was diluted with EtOAc (100 mL) and washed with H₂O (2×50 mL)and brine (50 mL) before being dried (MgSO₄), filtered and evaporatedunder reduced pressure. The crude product was purified by silica gelchromatography (gradient elution: 100% hexane to 7:3 v/v hexane/EtOAc)to afford pure product 12 (252.2 mg, 88% yield). Analytical data:ES⁺=1.95 min, m/z 657.25 [M+H]⁺

(b) tert-Butyl(11S,11aS)-2-(4-((S)-2-((S)-2-amino-3-methylbutanamido)propanamido)phenyl)-8-((5-(((11S,11aS)-10-(tert-butoxycarbonyl)-7-methoxy-2-methylene-5-oxo-11-((tetrahydro-2H-pyran-2-yl)oxy)-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-11-((tert-butyldimethylsilyl)oxy)-7-methoxy-5-oxo-11,11a-dihydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-10(5H)-carboxylate(13)

Monomer 10 (403 mg, 0.42 mmol, 1.1 eq.) and monomer 12 (250 mg, 0.38mmol, 1eq.) were solubilised in dry DMF (5 mL) under argon.K₂CO₃ (80.8mg, 0.38 mmol, 1 eq.) was added and the mixture heated to 60° C. Uponcompletion the mixture was diluted with EtOAc (100 mL) and washed withH₂O (2×50 mL) and brine (50 mL) before being dried (MgSO₄), filtered andevaporated under reduced pressure. The crude product was purified bysilica gel chromatography (gradient elution: 100% CHCl₃ to 9:1 v/vCHCl₃/MeOH) to afford pure product 13 (260.9 mg, 54% yield). Analyticaldata: ES⁺=1.60 min, m/z 1267.25 [M+H]⁺

(c) tert-Butyl(11S,11aS)-8-((5-(((11S,11aS)-10-(tert-butoxycarbonyl)-7-methoxy-2-methylene-5-oxo-11-((tetrahydro-2H-pyran-2-yl)oxy)-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-11-((tert-butyldimethylsilyl)oxy)-2-(4-((2S,5S)-37-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-5-isopropyl-2-methyl-4,7,35-trioxo-10,13,16,19,22,25,28,31-octaoxa-3,6,34-triazaheptatriacontanamido)phenyl)-7-methoxy-5-oxo-11,11a-dihydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-10(5H)-carboxylate(14)

EDCl hydrochloride (39 mg, 0.197 mmol, 1 eq.) was added to a suspensionof maleimide-PEG8-acid (117 mg, 0.197 mmol, 1 eq.) in dry CH₂Cl₂ (5 mL)under argon atmosphere. The mixture was stirred for 30 min at roomtemperature before PBD 13 (250 mg, 0.197 mmol) was added. Stirring wasmaintained until the reaction was complete (usually 5 hours). Thereaction was diluted with CH₂Cl₂ and the organic phase was washed withH₂O and brine before being dried over MgSO₄, filtered and excess solventremoved by rotary evaporation under reduced pressure. The product waspurified by careful silica gel chromatography (gradient elution: 100%CHCl₃ to 9:1 v/v CHCl₃/MeOH) to afford pure product 14 (273.8 mg, 75%yield). Analytical data: ES⁺=1.99 min, m/z 1863.95 [M+Na]⁺

(d)1-(3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanamido)-N-((2S)-1-(((2S)-1-((4-(7-methoxy-8-((5-((7-methoxy-2-methylene-5-oxo-2,3,5,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-5-oxo-5,11a-dihydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-3,6,9,12,15,18,21,24-octaoxaheptacosan-27-amide (15)

To PBD 14 (260 mg, 0.14 mmol, 1 eq.) was added H₂O (80 μL) and theslurry was cooled to 0° C. TFA (2 mL) was then added and the mixture wasleft to stir at 0° C. until completion. The reaction was subsequentlyquenched with aqueous NaHCO_(3 (sat.)) and extracted with CH₂Cl₂ (2×50mL). The combined organics were then washed with H₂O (50 mL) and brine(50 mL) before being dried (MgSO₄), filtered and evaporated underreduced pressure. The crude was purified by preparative HPLC and theproduct fractions combined and freeze dried on the freeze drierovernight to afford pure product 15 (29.9 mg, 16% yield). Analyticaldata: ES*=1.41 min, m/z 1407.25 [M+H]⁺

Example 2

Conjugation A

Trastuzumab (10 mg, 66.7 nanomoles) was diluted into 9.0 mL of areduction buffer containing 10 mM sodium borate pH 8.4, 2.5 mM EDTA anda final antibody concentration of 1.11 mg/mL. A 10 mM solution of TCEPwas added (1.7 molar equivalent/antibody, 113.3 nanomoles, 11.33 μL) andthe reduction mixture was heated at +37° C. for 1.5 hours in anincubator with 150 rpm shaking. After cooling down to room temperature,compound 14 was added as a DMSO solution (7.5 molar equivalent/antibody,525 nanomoles, in 1.0 mL DMSO). The solution was mixed for 1.5 hours atroom temperature, then the conjugation was quenched by addition ofN-acetyl cysteine (1 micromole, 100; AL at 10 mM), then injected into anAKTA™ Pure FPLC using a GE Healthcare HiLoad™ 26/600 column packed withSuperdex 200 PG, eluting with 2.6 mL/min of sterile-filteredphosphate-buffered saline (PBS). Fractions corresponding to ConjAmonomer peak were pooled, concentrated using a 15 mL Amicon Ultracell50KDa MWCO spin filter, analysed and sterile-filtered.

UHPLC analysis on a Shimadzu Prominence system using a Phenomenex Aeris3.6u XB-C18 150 mm×2.1 mm column eluting with a gradient of water andacetonitrile on a reduced sample of ConjA at 280 nm and 330 nm (Compound14 specific) shows a mixture of light and heavy chains attached toseveral molecules of compound 14, consistent with a drug-per-antibodyratio (DAR) of 2.59 molecules of compound 14 per antibody.

UHPLC analysis on a Shimadzu Prominence system using a Phenomenex Yarra3u SEC-3000 300 mm×4.60 mm eluting with sterile-filtered SEC buffercontaining 200 mM potassium phosphate pH 6.95, 250 mM potassium chlorideand 10% isopropanol (v/v) on a sample of ConjA at 280 nm shows a monomerpurity of over 97% with no impurity detected. UHPLC SEC analysis gives aconcentration of final ConjA at 1.59 mg/mL in 3.8 mL, obtained mass ofConjA is 6.04 mg (60% yield).

Conjugation B

A 50 mM solution of tris(2-carboxyethyl)phosphine hydrochloride (TCEP)in phosphate-buffered saline pH 7.4 (PBS) was added (50 molarequivalent/antibody, 35 micromoles, 700 μL) to a 24.14 mL solution ofantibody (105 mg, 700 nanomoles) in reduction buffer containing PBS and1 mM ethylenediaminetetraacetic acid (EDTA) and a final antibodyconcentration of 4.35 mg/mL. The reduction mixture was heated at +37° C.for 3 hours (or until full reduction is observed by UHPLC) in anincubator with gentle (<150 rpm) shaking. After cooling down to roomtemperature, the reduced antibody was buffer exchanged, via spin filtercentrifugation, into a reoxidation buffer containing PBS pH 7.4 and 1 mMEDTA to remove all the excess reducing agent. A 50 mM solution ofdehydroascorbic acid (DHAA, 10 molar equivalent/antibody, 7 micromoles,140 NL) in DMSO was added and the reoxidation mixture was allowed toreact for 16 hours at room temperature with gentle (<150 rpm) shaking atan antibody concentration of 2.3 mg/mL (or more DHAA added and reactionleft for longer until full reoxidation of the cysteine thiols to reformthe inter-chain cysteine disulfides is observed by UHPLC). Thereoxidation mixture was then sterile-filtered and diluted in aconjugation buffer containing PBS pH 7.4, 1 mM EDTA for a final antibodyconcentration of 1.0-1.5 mg/mL. Compound 14 was added as a DMSO solution(10 molar equivalent/antibody, 1 micromole, in 1.0 mL DMSO) to 9 mL ofthis reoxidised antibody solution (15 mg, 100 nanomoles) for a 10% (v/v)final DMSO concentration. The solution was mixed for 1.5 hours at roomtemperature, then the conjugation was quenched by addition of N-acetylcysteine (4 micromoles, 404 at 100 mM), then diluted to >50 mL in PBSand sterile-filtered, ready for tangential flow filtration (TFF)purification.

UHPLC analysis on a Shimadzu Prominence system using a Phenomenex Aeris3.6u XB-C18 150 mm×2.1 mm column eluting with a gradient of water andacetonitrile on a reduced sample of ConjB at 280 nm and 330 nm (Compound14 specific) shows unconjugated light chains and a mixture ofunconjugated heavy chains and heavy chains attached to a single moleculeof Compound 14, consistent with a drug-per-antibody ratio (DAR) of 1.88molecules of Compound 14 per antibody.

UHPLC analysis on a Shimadzu Prominence system using a Tosoh BioscienceTSKgel SuperSW mAb HTP 4 μm 4.6×150 mm column (with a 4 μm 3.0×20 mmguard column) eluting with 0.3 mL/minute sterile-filtered SEC buffercontaining 200 mM potassium phosphate pH 6.95, 250 mM potassium chlorideand 10% isopropanol (v/v) on a sample of ConjB at 280 nm shows a monomerpurity of 96%. UHPLC SEC analysis gives a concentration of final ConjBat 0.27 mg/mL in 50 mL, obtained mass of ConjB is 13.35 mg (89% yield).

Example 3

CB.17 SCID mice, aged 8-12 weeks, were injected with 1 mm³ BT474 tumourfragments subcutaneously in the flank. When tumours reach an averagesize of 100-150 mm³, treatment is begun. Mice are weighed twice a week.Tumour size is measured twice a week. Animals are monitoredindividually. The endpoint of the experiment is a tumour volume of 1000mm³ or 59 days, whichever comes first.

Groups of 10 xenografted mice were injected i.v. with 0.2 ml of antibodydrug conjugate (ADC), in phosphate buffered saline (vehicle) or with 0.2ml of vehicle alone. The concentration of ADC was adjusted to give 0.3or 1.0 mg ADC/kg body weight in a single dose.

FIG. 1 shows the effect on mean tumour volume in groups of 10 mice dosedwith ConjA at 0.3 or 1.0 mg/kg compared to vehicle control.

All regimens were acceptably tolerated with little body weight loss. Themedian time to end point (TTE) for vehicle-treated controls was 23.5days, establishing a maximum possible tumour growth delay (TGD) of 35.5days (151%) for the 59-day study. The ConjA regimen resulted in themaximum possible TGD, had ten of ten 59-day survivors and producedsurvival benefit that was statistically significantly different fromvehicle-treated controls (P<0.001).

ConjA, at 1 mg/kg produced 100% regressions, nine partial regressions(PRs) and 1 complete regression (CR) which remained a tumour-freesurvivor (TFS) at study end.

ConjA at 0.3 mg/kg produced 60% regressions consisting of five PRs andone CR, that remained a TFS at study end.

Example 4—Toxicity Studies/Therapeutic Index

A single dose NonGLP toxicity study was used to determine the maximumtolerated dose (MTD) and safety profile of ConjA. Male Sprague Dawleyrats (Harlan, Inc) were dosed once by slow bolus intravenous injectionvia the tail vein with vehicle control (25 mM Histidine-HCl, 7% sucrose,0.02% Polysorbate 80, pH 6.0) or ConjA. Parameters evaluated during thestudy included mortality, physical examinations, cageside observations,body weights, body weight changes, clinical pathology (clinicalchemistry, hematology, and coagulation), and gross pathology findings.

Animals that received 0.5 mg/kg of ConjA survived until schedulednecropsy on Study Day (SD) 29. Animals that received 1.0 mg/kg of ConjAwere either found dead, or euthanized due to morbidity by SD 8.

Male Rats Dose Dose Main Study Group Treatment Route (mg/kg) Frequency N1 Control IV 0 Single 5 3 ConjA IV 0.5 Single 5 6 ConjA IV 1.0 Single 5Control/Vehicle for dilution = 25 mM Histidine-HCl, 7% sucrose, 0.02%Polysorbate 80, pH 6.0

Tolerability was determined based on toxicity end points, including bodyweight loss (>15%) and bone marrow suppression. Based on a lack ofsevere toxicity at 0.5 mg/kg, and mortality at 1.0 mg/kg, the maximumtolerated dose (MTD) in the rat after a single dose of ConjA wasdetermined to be 0.5 mg/kg.

When compared to the minimum effective dose (MED) of 0.3 mg/kg in theBT474 xenograft model (see above), the potential therapeutic index (TI)for ConjA is 1.7 in rats, calculated as follows:TI=MTD in rat (mg/kg)/MED in mouse efficacy model (mg/kg)

All documents and other references mentioned above are hereinincorporated by reference.

The invention claimed is:
 1. A compound with the formula I:

wherein: R² is of formula II′:

where A is a C₅₋₇ aryl group, X is selected from the group consistingof: OH, SH, CO₂H, COH, N═C═O, NHNH₂, CONHNH₂,

 and NHR^(N), wherein R^(N) is selected from H and C₁₋₄ alkyl, andeither: (i) Q¹ is a single bond, and Q² is selected from a single bondand —Z—(CH₂)_(n)—, where Z is selected from a single bond, O, S and NHand n is from 1 to 3; or (ii) Q¹ is —CH═CH—, and Q² is a single bond;R¹² is selected from the group consisting of ═CH₂, ═CHR^(D1) and═CR^(D1)R^(D2), where R^(D1) and R^(D2) are independently selected fromR, CO₂R, COR, CHO, CO₂H, and halo; R⁶ and R⁹ are independently selectedfrom the group consisting of: H, R, OH, OR, SH, SR, NH₂, NHR, NRR′,nitro, Me₃Sn and halo; R⁷ is selected from the group consisting of: H,R, OH, OR, SH, SR, NH₂, NHR, NHRR′, nitro, Me₃Sn and halo; where R andR′ are independently selected from optionally substituted C₁₋₁₂ alkyl,C₃₋₂₀ heterocyclyl and C₅₋₂₀ aryl groups; either: (a) R¹⁰ is H, and R¹¹is OH, OR^(A), where R^(A) is C₁₋₄ alkyl; (b) R¹⁰ and R¹¹ form anitrogen-carbon double bond between the nitrogen and carbon atoms towhich they are bound; or (c) R¹⁰ is H and R¹¹ is SO_(z)M, where z is 2or 3 and M is a monovalent pharmaceutically acceptable cation; R″ is aC₃₋₁₂ alkylene group, which chain is optionally interrupted by one ormore heteroatoms, and/or aromatic rings; Y and Y′ are independentlyselected from the group consisting of O, S, and NH; R^(6′), R^(7′),R^(9′) are selected from the same groups as R⁶, R⁷ and R⁹ respectivelyand R^(10′) and R^(11′) are the same as R¹⁰ and R¹¹, wherein if R¹¹ andR^(11′) are SO_(Z)M, M may represent a divalent pharmaceuticallyacceptable cation.
 2. A compound according to claim 1 wherein Y and Y′are O, and R″ is unsubstituted C₃₋₇ alkylene.
 3. A compound according toclaim 1, wherein R⁶ and R⁹ are H.
 4. A compound according to claim 1,wherein R⁷ is selected from C₁₋₄ alkyoxy and benzyloxy.
 5. A compoundaccording to claim 1, wherein R² is phenyl.
 6. A compound according toclaim 1, wherein X is NH₂.
 7. A compound according to claim 1, whereinR¹² is ═CH₂.
 8. A compound according to claim 1, which is of formula Ia:

where R^(1a) is selected from Me and Ph, and the amino group is ateither the meta or para positions of the phenyl group.
 9. A compound offormula II:

wherein R², R¹², R⁶, R⁷, R⁹, Y, R″, Y′, R^(6′), R^(7′) and R^(9′) are asdefined in claim 1, and either: (a) R¹⁰ is carbamate nitrogen protectinggroup, and R¹¹ is O-Prot^(o), wherein Prot^(o) is an oxygen protectinggroup; or (b) R¹⁰ is a hemi-aminal nitrogen protecting group and R¹¹ isan oxo group.
 10. A conjugate of formula (111a-1):L-(A¹-L¹-D)_(p)  (IIIa-1) wherein A¹ is of formula:-L^(A)-A²- wherein L^(A) is selected from:

where Ar represents a C₅₋₆ arylene group, and A² is selected from:

where n is 0 to 6;

where n is 0 to 6;

where n is 0 or 1, and m is 0 to 30;

where n is 0 or 1, and m is 0 to 30; where the asterisk indicates thepoint of attachment to L¹, the wavy line indicates the point ofattachment to L^(A); and D is a Drug unit which is a PBD dimer accordingto claim 1, wherein L¹ is connected to D via the X substituent of R²,and p is an integer of from 1 to
 20. 11. A conjugate of claim 10,wherein L^(A) is (L^(A1-1)):

and A² is

where n is 1, and m is 0 to
 8. 12. A conjugate of claim 10, wherein L¹is a dipeptide.
 13. The Conjugate of claim 12, wherein L¹ is selectedfrom the group consisting of valine-alanine, valine-citrulline andphenyalanine-lysine.
 14. A drug linker of formula IV:DLU-D  (IV) wherein DLU is a Drug Linker unit, and D is a Drug unitwhich is a PBD dimer according to any claim 1, wherein DLU is connectedto D via the X substituent of R²; wherein DLU is of the formula:G¹-L¹-, wherein G¹ is of formula:G^(A)-A²- where G^(A) is selected from:

where Ar represents a C₅₋₆ arylene group, and A² is selected from:

where n is 0 to 6;

where n is 0 to 6;

where n is 0 or 1, and m is 0 to 30;

where n is 0 or 1, and m is 0 to 30; where the asterisk indicates thepoint of attachment to L¹, the wavy line indicates the point ofattachment to G^(A).
 15. A drug linker of claim 14, wherein G^(A) is(G^(A1-1)):

and A² is

where n is 1, and m is 0 to
 8. 16. A drug linker of claim 15, wherein mis
 8. 17. A drug linker of claim 14, wherein L¹ is a dipeptide.
 18. Adrug linker of claim 17, wherein L¹ is selected from the groupconsisting of valine-alanine, valine-citrulline and phenyalanine-lysine.19. A method of treatment of a proliferative disease comprisingadministering to a subject in need comprising administering to a subjectin need of treatment a therapeutically effective amount of a compoundaccording to claim
 1. 20. A method of treatment of a proliferativedisease comprising administering to a subject in need comprisingadministering to a subject in need of treatment a therapeuticallyeffective amount of a conjugate according to claim 10.